Metalloenzyme inhibitor compounds

ABSTRACT

The instant invention describes compounds having metalloenzyme modulating activity, and methods of treating diseases, disorders or symptoms thereof mediated by such metalloenzymes.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application claims priority benefit under 35 U.S.C. §119(e)of U.S. Provisional Patent Application Ser. No. 61/500,372, filed Jun.23, 2011; and U.S. Provisional Patent Application Ser. No. 61/611,917,filed Mar. 16, 2012. The disclosures of these applications areincorporated herein by reference.

BACKGROUND

Living organisms have developed tightly regulated processes thatspecifically import metals, transport them to intracellular storagesites and ultimately transport them to sites of use. One of the mostimportant functions of metals such as zinc and iron in biologicalsystems is to enable the activity of metalloenzymes. Metalloenzymes areenzymes that incorporate metal ions into the enzyme active site andutilize the metal as a part of the catalytic process. More thanone-third of all characterized enzymes are metalloenzymes.

The function of metalloenzymes is highly dependent on the presence ofthe metal ion in the active site of the enzyme. It is well recognizedthat agents which bind to and inactivate the active site metal iondramatically decrease the activity of the enzyme. Nature employs thissame strategy to decrease the activity of certain metalloenzymes duringperiods in which the enzymatic activity is undesirable. For example, theprotein TIMP (tissue inhibitor of metalloproteases) binds to the zincion in the active site of various matrix metalloprotease enzymes andthereby arrests the enzymatic activity. The pharmaceutical industry hasused the same strategy in the design of therapeutic agents. For example,the azole antifungal agents fluconazole and voriconazole contain a1-(1,2,4-triazole) group that binds to the heme iron present in theactive site of the target enzyme lanosterol demethylase and therebyinactivates the enzyme. Another example includes the zinc-bindinghydroxamic acid group that has been incorporated into most publishedinhibitors of matrix metalloproteinases and histone deacetylases.Another example is the zinc-binding carboxylic acid group that has beenincorporated into most published angiotensin-converting enzymeinhibitors.

In the design of clinically safe and effective metalloenzyme inhibitors,use of the most appropriate metal-binding group for the particulartarget and clinical indication is critical. If a weakly bindingmetal-binding group is utilized, potency may be suboptimal. On the otherhand, if a very tightly binding metal-binding group is utilized,selectivity for the target enzyme versus related metalloenzymes may besuboptimal. The lack of optimal selectivity can be a cause for clinicaltoxicity due to unintended inhibition of these off-targetmetalloenzymes. One example of such clinical toxicity is the unintendedinhibition of human drug metabolizing enzymes such as cytochrome P4502C9 (CYP2C9), CYP2C19 and CYP3A4 by the currently-available azoleantifungal agents such as fluconazole and voriconazole. It is believedthat this off-target inhibition is caused primarily by theindiscriminate binding of the currently utilized 1-(1,2,4-triazole) toiron in the active site of CYP2C9, CYP2C19 and CYP3A4. Another exampleof this is the joint pain that has been observed in many clinical trialsof matrix metalloproteinase inhibitors. This toxicity is considered tobe related to inhibition of off-target metalloenzymes due toindiscriminate binding of the hydroxamic acid group to zinc in theoff-target active sites.

Therefore, the search for metal-binding groups that can achieve a betterbalance of potency and selectivity remains an important goal and wouldbe significant in the realization of therapeutic agents and methods toaddress currently unmet needs in treating and preventing diseases,disorders and symptoms thereof.

Fungicides are compounds, of natural or synthetic origin, which act toprotect and cure plants against damage caused by agriculturally relevantfungi. Generally, no single fungicide is useful in all situations.Consequently, research is ongoing to produce fungicides that may havebetter performance, are easier to use, and cost less.

The present disclosure relates to compounds of Formula I, shown below,and their derivatives and their use as fungicides. The compounds of thepresent disclosure may offer protection against ascomycetes,basidiomycetes, deuteromycetes and oomycetes.

BRIEF SUMMARY OF THE INVENTION

The invention is directed towards compounds (e.g., any of thosedelineated herein), methods of modulating activity of metalloenzymes,and methods of treating diseases, disorders or symptoms thereof. Themethods can comprise the compounds herein.

A method of controlling a pathogen-induced disease in a plant that is atrisk of being diseased from the pathogen comprising contacting one ofthe plant and an area adjacent to the plant with a composition ofFormula I, or salt, solvate, hydrate or prodrug thereof, wherein:

MBG is optionally substituted tetrazolyl, optionally substitutedtriazolyl, optionally substituted oxazolyl, optionally substitutedpyrimidinyl, optionally substituted thiazolyl, or optionally substitutedpyrazolyl;

R₁ is H, halo, alkyl or haloalkyl;

R₂ is H, halo, alkyl or haloalkyl;

R₃ is 1,1′-biphenyl substituted with 4′-OCH₂CF₃ or 4′-F, or heteroaryl,which may be optionally substituted with 1, 2 or 3 independent R₅;

R₄ is aryl, heteroaryl, alkyl or cycloalkyl, optionally substituted with0, 1, 2 or 3 independent R₆;

each R₅ is independently H, halo, aryl optionally substituted with 1, 2or 3 independent R₆, heteroaryl, haloalkyl, haloalkoxy, cyano, nitro,alkyl, alkoxy, alkenyl, haloalkenyl, arylalkenyl, alkynyl, haloalkynyl,alkylaryl, arylalkynyl, arylalkyl, cycloalkyl, halocycloalkyl,thioalkyl, SF₃, SF₆, SCN, SO₂R₇, C(O)alkyl, C(O)OH, C(O)Oalkyl;

each R₆ is independently alkyl, thioalkyl, cyano, haloalkyl, hydroxy,alkoxy, halo, haloalkoxy, —C(O)alkyl, —C(O)OH, —C(O)Oalkyl, SF₃, SF₆,SCN, SO₃H; and SO₂R₇;

R₇ is independently alkyl, aryl, substituted aryl, heteroaryl orsubstituted heteroaryl;

R₈ is H, —Si(R₉)₃, —P(O)(OH)₂, —CH₂—O—P(O)(OH)₂, or —C(O)alkyloptionally substituted with amino;

R₉ is independently alkyl or aryl;

and wherein R₃ is not 2-pyridyl optionally substituted with 1, 2 or 3independent R₅.

-   -   Other aspects are a compound of the formulae herein:    -   wherein R₁ is fluoro;    -   wherein R₂ is fluoro;    -   wherein R₁ and R₂ are fluoro;    -   wherein R₄ is phenyl optionally substituted with 0, 1, 2 or 3        independent R₆;    -   wherein R₄ is phenyl optionally substituted with 0, 1, 2 or 3        independent halo;    -   wherein R₄ is phenyl optionally substituted with 0, 1, 2 or 3        independent fluoro;    -   wherein R₄ is 2,4-difluorophenyl;    -   wherein R₅ is halo;    -   wherein R₃ is heteroaryl other than 2-pyridyl, optionally        substituted with 1, 2 or 3 independent R₅;    -   wherein at least one R₅ is halo;    -   wherein:        -   R₁ is fluoro;        -   R₂ is fluoro;        -   R₄ is 2,4-difluorophenyl; and        -   R₃ is heteroaryl other than 2-pyridyl, substituted with 1, 2            or 3 independent R₅;    -   wherein:        -   R₁ is fluoro;        -   R₂ is fluoro;        -   R₄ is 2,4-difluorophenyl; and        -   R₃ is bicyclic heteroaryl substituted with 1, 2 or 3            independent R₅;    -   wherein R₃ is 2-quinolinyl substituted with 1, 2 or 3        independent R₅;    -   wherein MBG is optionally substituted tetrazolyl or optionally        substituted triazolyl;    -   wherein MBG is 1H-tetrazol-1-yl, 2H-tetrazol-2-yl,        4H-1,2,4-triazol-4-yl, or 1H-1,2,4-triazol-1-yl;    -   wherein MBG is 1H-tetrazol-1-yl, or 2H-tetrazol-2-yl;    -   wherein MBG is 4H-1,2,4-triazol-4-yl, or 1H-1,2,4-triazol-1-yl;    -   wherein R₃ is thienyl, thiazolyl, quinolinyl, pyridyl,        benzothiazolyl, pyrimidinyl, quinoxalinyl, pyrazinyl, or        pyridiazinyl, each optionally substituted with 1, 2 or 3        independent R₅;    -   wherein:        -   R₁ is fluoro;        -   R₂ is fluoro;        -   R₄ is 2,4-difluorophenyl; and        -   R₃ is thienyl, thiazolyl, quinolinyl, pyridyl,            benzothiazolyl, pyrimidinyl, quinoxalinyl, pyrazinyl, or            pyridiazinyl, each optionally substituted with 1, 2 or 3            independent R₅;    -   wherein R₃ is thienyl, thiazolyl, quinolinyl, pyridyl,        benzothiazolyl, pyrimidinyl, quinoxalinyl, pyrazinyl, or        pyridiazinyl, each optionally substituted with 1, 2 or 3        independent alkyl, alkenyl, alkoxy, halo, cyano, haloalkyl,        haloalkoxy, alkyl substituted with halophenyl, alkynyl        substituted with halophenyl, or phenyl substituted with        haloalkyl, haloalkoxy, halo, or cyano.

The compounds herein include those wherein the compound is identified asattaining affinity, at least in part, for a metalloenzyme by formationof one or more of the following types of chemical interactions or bondsto a metal: sigma bonds, covalent bonds, coordinate-covalent bonds,ionic bonds, pi bonds, delta bonds, or backbonding interactions. Thecompounds can also attain affinity through weaker interactions with themetal such as van der Waals interactions, pi-cation interactions,pi-anion interactions, dipole-dipole interactions, ion-dipoleinteractions. In one aspect, the compound is identified as having abonding interaction with the metal via the 1-tetrazolyl moiety; inanother aspect, the compound is identified as having a bondinginteraction with the metal via the N2 of the 1-tetrazolyl moiety; inanother aspect, the compound is identified as having a bondinginteraction with the metal via the N3 of the 1-tetrazolyl moiety; inanother aspect, the compound is identified as having a bondinginteraction with the metal via the N4 of the 1-tetrazolyl moiety. In oneaspect, the compound is identified as having a bonding interaction withthe metal via the 4-triazolyl moiety; in another aspect, the compound isidentified as having a bonding interaction with the metal via the N1 ofthe 4-triazolyl moiety; in another aspect, the compound is identified ashaving a bonding interaction with the metal via the N2 of the4-tetrazolyl moiety.

Methods for assessing metal-ligand binding interactions are known in theart as exemplified in references including, for example, “Principles ofBioinorganic Chemistry” by Lippard and Berg, University Science Books,(1994); “Mechanisms of Inorganic Reactions” by Basolo and Pearson, JohnWiley & Sons Inc; 2nd edition (September 1967); “Biological InorganicChemistry” by Ivano Bertini, Harry Gray, Ed Stiefel, Joan Valentine,University Science Books (2007); Xue et al. “Nature Chemical Biology”,vol. 4, no. 2, 107-109 (2008).

In certain instances, the compounds of the invention are selected fromthe following of Formula I (and pharmaceutically and agriculturallyacceptable salts, solvates, or hydrates thereof)

-   1-(5-Chlorothiophen-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (1);-   1-(4-Bromothiazol-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (2);-   4-(2-(2-(2,4-Difluorophenyl)-1,1-difluoro-2-hydroxy-3-(1H-tetrazol-1-yl)propyl)thiazol-4-yl)benzonitrile    (3);-   1-(6-Chloroquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (4);-   2-(2,4-Difluorophenyl)-1,1-difluoro-1-(quinolin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol    (5);-   1-(Benzo[d]thiazol-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (6);-   2-(2,4-Difluorophenyl)-1,1-difluoro-1-(pyrimidin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol    (7);-   2-(4-Chloro-2-fluorophenyl)-1-(6-chloroquinolin-2-yl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (8);-   1-(6-Bromoquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (9);-   1-(6-Chloroquinoxalin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (10);-   1-(6-Chlorobenzo[d]thiazol-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (11);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(thiazol-2-yl)propan-2-ol    (12);-   1-(5-Bromothiophen-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (13);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(thiophen-2-yl)propan-2-ol    (14);-   1-(6-Chloroquinolin-2-yl)-1,1-difluoro-2-(4-methoxyphenyl)-3-(1H-tetrazol-1-yl)propan-2-ol    (15);-   1-(6-Chloroquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(2H-tetrazol-2-yl)propan-2-ol    (16);-   2-(2,4-Difluorophenyl)-1,1-difluoro-1-(6-fluoroquinolin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol    (17);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-(trifluoromethyl)quinolin-2-yl)propan-2-ol    (18);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-(2,2,2-trifluoroethoxy)quinolin-2-yl)propan-2-ol    (19);-   1-(6-Chloroquinolin-2-yl)-1,1-difluoro-2-(2-fluoro-4-(trifluoromethyl)phenyl)-3-(1H-tetrazol-1-yl)propan-2-ol    (20);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-(trifluoromethoxy)quinolin-2-yl)propan-2-ol    (21);-   2-(2-Chloro-4-(trifluoromethyl)phenyl)-1-(6-chloroquinolin-2-yl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (22);-   1-(6-Chloroquinolin-2-yl)-2-(3,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (23);-   2-(2-(2,4-Difluorophenyl)-1,1-difluoro-2-hydroxy-3-(1H-tetrazol-1-yl)propyl)quinoline-6-carbonitrile    (24);-   1-(6-(Difluoromethyl)quinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (25);-   2-(2,4-Difluorophenyl)-1,1-difluoro-1-(6-methylquinolin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol    (26);-   1-(6-Bromobenzo[d]thiazol-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (27);-   1-(6-Chloroquinolin-2-yl)-2-(2,5-difluorophenyl)-1,1-difluoro-3-(2H-tetrazol-2-yl)propan-2-ol    (28);-   1-(5,6-Dichloroquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (29);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(2,2,2-trifluoroethoxy)quinolin-2-yl)propan-2-ol    (30);-   1-(5-Chloroquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (31);-   1-(6-Chloroquinolin-2-yl)-1,1-difluoro-2-(4-fluorophenyl)-3-(1H-tetrazol-1-yl)propan-2-ol    (32);-   1-(6-Cloroquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-1,2,4-triazol-1-yl)propan-2-ol    (33);-   2-(4-Chloro-2-fluorophenyl)-1-(6-chloroquinoxalin-2-yl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (34);-   1-(6-Chloroquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(4H-1,2,4-triazol-4-yl)propan-2-ol    (35);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-3-yl)propan-2-ol    (36);-   1-(7-Chloroisoquinolin-3-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (37);-   1-(6-Bromoquinoxalin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (38);-   1-(5-(4-(Difluoromethoxy)phenyl)pyrazin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (39);-   1-(5-(4-Chlorophenyl)pyrazin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (40);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyrazin-2-yl)propan-2-ol    (41);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(trifluoromethoxy)phenyl)pyrazin-2-yl)propan-2-ol    (42);-   1-(5-(4-Bromophenyl)pyrazin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (43);-   2-(2,4-Difluorophenyl)-1-(5-(3,4-difluorophenyl)pyrazin-2-yl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (44);-   1,1-Difluoro-2-(4-fluorophenyl)-3-(1H-tetrazol-1-yl)-1-(5-(4-(trifluoromethoxy)phenyl)pyrazin-2-yl)propan-2-ol    (45);-   2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(4-fluorophenyl)pyrazin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol    (46);-   2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(4-fluorophenyl)pyrazin-2-yl)-3-(2H-tetrazol-2-yl)propan-2-ol    (47);-   2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(4-methoxyphenyl)pyrazin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol    (48);-   1-(5-Chloropyrazin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol    (49);-   2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-((4-fluorophenyl)ethynyl)pyrazin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol    (50);-   2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-((4-fluorophenyl)ethynyl)pyrazin-2-yl)-3-(2H-tetrazol-2-yl)propan-2-ol    (51);-   2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(4-fluorophenethyl)pyrazin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol    (52);-   2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(4-fluorophenethyl)pyrazin-2-yl)-3-(2H-tetrazol-2-yl)propan-2-ol    (53);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-(trifluoromethoxy)quinoxalin-2-yl)propan-2-ol    (54);-   2-(2,4-Difluorophenyl)-1,1-difluoro-1-(6-fluoroquinoxalin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol    (55);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-(4-(trifluoromethyl)phenyl)pyridazin-3-yl)propan-2-ol    (56);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-(4-(trifluoromethoxy)phenyl)pyridazin-3-yl)propan-2-ol    (57);-   2-(2,4-Difluorophenyl)-1,1-difluoro-1-(6-(4-fluorophenyl)pyridazin-3-yl)-3-(1H-tetrazol-1-yl)propan-2-ol    (58);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-vinylquinoxalin-2-yl)propan-2-ol    (59);-   2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(4′-(2,2,2-trifluoroethoxy)-[1,1′-biphenyl]-4-yl)propan-2-ol    (60);-   2-(2,4-Difluorophenyl)-1,1-difluoro-1-(4′-fluoro-[1,1′-biphenyl]-4-yl)-3-(1H-tetrazol-1-yl)propan-2-ol    (61).

In another aspect, the invention provides an agricultural compositioncomprising the compound of Formula I and an agriculturally acceptablecarrier.

In other aspects, the invention provides a compound of any of theformulae herein, wherein the compound inhibits (or is identified toinhibit) lanosterol demethylase (CYP51).

In other aspects, the invention provides a compound of any of theformulae herein, wherein the compound is identified as having anactivity range against a target organism (e.g., C. albicans minimuminhibitory concentration (MIC)<0.25 micrograms per milliliter (μg/mL);S. tritici minimum inhibitory concentration (MIC)<0.5 micrograms permilliliter (μg/mL); e.g., P. triticina minimum inhibitory concentration(MIC)<0.5 micrograms per milliliter (μg/mL).

In another aspect, the invention provides a pharmaceutical compositioncomprising the compound of Formula I and a pharmaceutically acceptablecarrier.

In other aspects, the invention provides a method of modulatingmetalloenzyme activity in a subject, comprising contacting the subjectwith a compound of Formula I, in an amount and under conditionssufficient to modulate metalloenzyme activity.

In one aspect, the invention provides a method of treating a subjectsuffering from or susceptible to a metalloenzyme-related disorder ordisease, comprising administering to the subject an effective amount ofa compound or pharmaceutical composition of Formula I.

In another aspect, the invention provides a method of treating a subjectsuffering from or susceptible to a metalloenzyme-related disorder ordisease, wherein the subject has been identified as in need of treatmentfor a metalloenzyme-related disorder or disease, comprisingadministering to said subject in need thereof, an effective amount of acompound or pharmaceutical composition of Formula I, such that saidsubject is treated for said disorder.

In another aspect, the invention provides a method of treating a subjectsuffering from or susceptible to a metalloenzyme-mediated disorder ordisease, wherein the subject has been identified as in need of treatmentfor a metalloenzyme-mediated disorder or disease, comprisingadministering to said subject in need thereof, an effective amount of acompound or pharmaceutical composition of Formula I, such thatmetalloenzyme activity in said subject is modulated (e.g., downregulated, inhibited).

The methods herein include those wherein the disease or disorder ismediated by any of 4-hydroxyphenyl pyruvate dioxygenase, 5-lipoxygenase,adenosine deaminase, alcohol dehydrogenase, aminopeptidase N,angiotensin converting enzyme, aromatase (CYP19), calcineurin, carbamoylphosphate synthetase, carbonic anhydrase family, catechol-O-methyltransferase, cyclooxygenase family, dihydropyrimidine dehydrogenase-1,DNA polymerase, farnesyl diphosphate synthase, farnesyl transferase,fumarate reductase, GABA aminotransferase, HIF-prolyl hydroxylase,histone deacetylase family, HIV integrase, HIV-1 reverse transcriptase,isoleucine tRNA ligase, lanosterol demethylase (CYP51), matrixmetalloprotease family, methionine aminopeptidase, neutralendopeptidase, nitric oxide synthase family, phosphodiesterase III,phosphodiesteraseIV, phosphodiesteraseV, pyruvate ferredoxinoxidoreductase, renal peptidase, ribonucleoside diphosphate reductase,thromboxane synthase (CYP5a), thyroid peroxidase, tyrosinase, urease, orxanthine oxidase.

The methods herein include those wherein the disease or disorder ismediated by any of 1-deoxy-D-xylulose-5-phosphate reductoisomerase(DXR), 17-alpha hydroxylase (CYP17), aldosterone synthase (CYP11B2),aminopeptidase P, anthrax lethal factor, arginase, beta-lactamase,cytochrome P450 2A6, D-Ala D-Ala ligase, dopamine beta-hydroxylase,endothelin converting enzyme-1, glutamate carboxypeptidase II,glutaminyl cyclase, glyoxalase, heme oxygenase, HPV/HSV E1 helicase,indoleamine 2,3-dioxygenase, leukotriene A4 hydrolase, methionineaminopeptidase 2, peptide deformylase, phosphodiesteraseVII, relaxase,retinoic acid hydroxylase (CYP26), TNF-alpha converting enzyme (TACE),UDP-(3-O-(R-3-hydroxymyristoyl))-N-acetylglucosamine deacetylase (LpxC),vascular adhesion protein-1 (VAP-1), or vitamin D hydroxylase (CYP24).

The methods herein include those wherein the disease or disorder iscancer, cardiovascular disease, inflammatory disease, infectiousdisease, metabolic disease, ophthalmologic disease, central nervoussystem (CNS) disease, urologic disease, or gastrointestinal disease.

The methods herein include those wherein the disease or disorder isprostate cancer, breast cancer, inflammatory bowel disease, psoriasis,systemic fungal infection, skin structure fungal infection, mucosalfungal infection, or onychomycosis.

Methods delineated herein include those wherein the subject isidentified as in need of a particular stated treatment. Identifying asubject in need of such treatment can be in the judgment of a subject ora health care professional and can be subjective (e.g. opinion) orobjective (e.g. measurable by a test or diagnostic method).

Another aspect of the invention is a composition comprising a compoundof a formulae herein (e.g., Formula I) and an agriculturally acceptablecarrier.

Another aspect of the invention is a method of treating or preventing ametalloenzyme-mediated disease or disorder in or on a plant comprisingcontacting a compound herein with the plant.

Another aspect of the invention is a method of inhibiting metalloenzymeactivity in or on a plant comprising contacting a compound herein withthe plant.

DETAILED DESCRIPTION

Definitions

In order that the invention may be more readily understood, certainterms are first defined here for convenience.

As used herein, the term “treating” a disorder encompasses preventing,ameliorating, mitigating and/or managing the disorder and/or conditionsthat may cause the disorder. The terms “treating” and “treatment” referto a method of alleviating or abating a disease and/or its attendantsymptoms. In accordance with the present invention “treating” includespreventing, blocking, inhibiting, attenuating, protecting against,modulating, reversing the effects of and reducing the occurrence ofe.g., the harmful effects of a disorder.

As used herein, “inhibiting” encompasses preventing, reducing andhalting progression. Note that “enzyme inhibition” (e.g., metalloenzymeinhibition) is distinguished and described below.

The term “modulate” refers to increases or decreases in the activity ofan enzyme in response to exposure to a compound of the invention.

The terms “isolated,” “purified,” or “biologically pure” refer tomaterial that is substantially or essentially free from components thatnormally accompany it as found in its native state. Purity andhomogeneity are typically determined using analytical chemistrytechniques such as polyacrylamide gel electrophoresis or highperformance liquid chromatography. Particularly, in embodiments thecompound is at least 85% pure, more preferably at least 90% pure, morepreferably at least 95% pure, and most preferably at least 99% pure.

The term “administration” or “administering” includes routes ofintroducing the compound(s) to a subject to perform their intendedfunction. Examples of routes of administration which can be used includeinjection (subcutaneous, intravenous, parenterally, intraperitoneally,intrathecal), topical, oral, inhalation, rectal and transdermal.

The term “effective amount” includes an amount effective, at dosages andfor periods of time necessary, to achieve the desired result. Aneffective amount of compound may vary according to factors such as thedisease state, age, and weight of the subject, and the ability of thecompound to elicit a desired response in the subject. Dosage regimensmay be adjusted to provide the optimum therapeutic response. Aneffective amount is also one in which any toxic or detrimental effects(e.g., side effects) of the inhibitor compound are outweighed by thetherapeutically beneficial effects.

The phrases “systemic administration,” “administered systemically”,“peripheral administration” and “administered peripherally” as usedherein mean the administration of a compound(s), drug or other material,such that it enters the patient's system and, thus, is subject tometabolism and other like processes.

The term “therapeutically or agriculturally effective amount” refers tothat amount of the compound being administered sufficient to preventdevelopment of or alleviate to some extent one or more of the symptomsof the condition or disorder being treated.

A therapeutically effective amount of compound (i.e., an effectivedosage) may range from about 0.005 micrograms per kilogram (μg/kg) toabout 200 milligrams per kilogram (mg/kg), preferably about 0.01 mg/kgto about 200 mg/kg, more preferably about 0.015 mg/kg to about 30 mg/kgof body weight. In other embodiments, the therapeutically effect amountmay range from about 1.0 picomolar (pM) to about 10 micromolar (μM). Theskilled artisan will appreciate that certain factors may influence thedosage required to effectively treat a subject, including but notlimited to the severity of the disease or disorder, previous treatments,the general health and/or age of the subject, and other diseasespresent. Moreover, treatment of a subject with a therapeuticallyeffective amount of a compound can include a single treatment or,preferably, can include a series of treatments. In one example, asubject is treated with a compound in the range of between about 0.005μg/kg to about 200 mg/kg of body weight, one time per day for betweenabout 1 to 10 weeks, preferably between about 2 to 8 weeks, morepreferably between about 3 to 7 weeks, and even more preferably forabout 4, 5, or 6 weeks. In another example, a subject may be treateddaily for several years in the setting of a chronic condition orillness. It will also be appreciated that the effective dosage of acompound used for treatment may increase or decrease over the course ofa particular treatment.

The term “chiral” refers to molecules which have the property ofnon-superimposability of the mirror image partner, while the term“achiral” refers to molecules which are superimposable on their mirrorimage partner.

The term “diastereomers” refers to stereoisomers with two or morecenters of dissymmetry and whose molecules are not minor images of oneanother.

The term “enantiomers” refers to two stereoisomers of a compound whichare non-superimposable minor images of one another. An equimolar mixtureof two enantiomers is called a “racemic mixture” or a “racemate.”

The term “isomers” or “stereoisomers” refers to compounds which haveidentical chemical constitution, but differ with regard to thearrangement of the atoms or groups in space.

The term “prodrug” includes compounds with moieties which can bemetabolized in vivo. Generally, the prodrugs are metabolized in vivo byesterases or by other mechanisms to active drugs. Examples of prodrugsand their uses are well known in the art (See, e.g., Berge et al. (1977)“Pharmaceutical Salts”, J. Pharm. Sci. 66:1-19). The prodrugs can beprepared in situ during the final isolation and purification of thecompounds, or by separately reacting the purified compound in its freeacid form or hydroxyl with a suitable esterifying agent. Hydroxyl groupscan be converted into esters via treatment with a carboxylic acid.Examples of prodrug moieties include substituted and unsubstituted,branched or unbranched lower alkyl ester moieties, (e.g., propionic acidesters), lower alkenyl esters, di-lower alkyl-amino lower-alkyl esters(e.g., dimethylaminoethyl ester), acylamino lower alkyl esters (e.g.,acetyloxymethyl ester), acyloxy lower alkyl esters (e.g.,pivaloyloxymethyl ester), aryl esters (phenyl ester), aryl-lower alkylesters (e.g., benzyl ester), substituted (e.g., with methyl, halo, ormethoxy substituents) aryl and aryl-lower alkyl esters, amides,lower-alkyl amides, di-lower alkyl amides, and hydroxy amides. Preferredprodrug moieties are propionic acid esters and acyl esters. Prodrugswhich are converted to active forms through other mechanisms in vivo arealso included. In aspects, the compounds of the invention are prodrugsof any of the formulae herein.

The term “subject” refers to animals such as mammals, including, but notlimited to, primates (e.g., humans), cows, sheep, goats, horses, dogs,cats, rabbits, rats, mice and the like. In certain embodiments, thesubject is a human.

The terms “a,” “an,” and “the” refer to “one or more” when used in thisapplication, including the claims. Thus, for example, reference to “asample” includes a plurality of samples, unless the context clearly isto the contrary (e.g., a plurality of samples), and so forth.

Throughout this specification and the claims, the words “comprise,”“comprises,” and “comprising” are used in a non-exclusive sense, exceptwhere the context requires otherwise.

As used herein, the term “about,” when referring to a value is meant toencompass variations of, in some embodiments ±20%, in some embodiments±10%, in some embodiments ±5%, in some embodiments ±1%, in someembodiments ±0.5%, and in some embodiments ±0.1% from the specifiedamount, as such variations are appropriate to perform the disclosedmethods or employ the disclosed compositions.

Use of the word “inhibitor” herein is meant to mean a molecule thatexhibits activity for inhibiting a metalloenzyme. By “inhibit” herein ismeant to decrease the activity of a metalloenzyme, as compared to theactivity of a metalloenzyme in the absence of the inhibitor. In someembodiments, the term “inhibit” means a decrease in metalloenzymeactivity of at least about 5%, at least about 10%, at least about 20%,at least about 25%, at least about 50%, at least about 60%, at leastabout 70%, at least about 80%, at least about 90%, or at least about95%. In other embodiments, inhibit means a decrease in metalloenzymeactivity of about 5% to about 25%, about 25% to about 50%, about 50% toabout 75%, or about 75% to 100%. In some embodiments, inhibit means adecrease in metalloenzyme activity of about 95% to 100%, e.g., adecrease in activity of 95%, 96%, 97%, 98%, 99%, or 100%. Such decreasescan be measured using a variety of techniques that would be recognizableby one of skill in the art. Particular assays for measuring individualactivity are described below.

Furthermore the compounds of the invention include olefins having eithergeometry: “Z” refers to what is referred to as a “cis” (same side)configuration whereas “E” refers to what is referred to as a “trans”(opposite side) configuration. With respect to the nomenclature of achiral center, the terms “d” and “l” configuration are as defined by theIUPAC Recommendations. As to the use of the terms, diastereomer,racemate, epimer and enantiomer, these will be used in their normalcontext to describe the stereochemistry of preparations.

As used throughout this specification, the term ‘R’ refers to the groupconsisting of C₁₋₈ alkyl, C₃₋₈ alkenyl or C₃₋₈ alkynyl, unless statedotherwise.

As used herein, the term “alkyl” refers to a straight-chained orbranched hydrocarbon group containing 1 to 12 carbon atoms. The term“lower alkyl” refers to a C₁-C₆ alkyl chain. Examples of alkyl groupsinclude methyl, ethyl, n-propyl, isopropyl, tert-butyl, and n-pentyl.Alkyl groups may be optionally substituted with one or moresubstituents.

The term “alkenyl” refers to an unsaturated hydrocarbon chain that maybe a straight chain or branched chain, containing 2 to 12 carbon atomsand at least one carbon-carbon double bond. Alkenyl groups may beoptionally substituted with one or more substituents.

The term “alkynyl” refers to an unsaturated hydrocarbon chain that maybe a straight chain or branched chain, containing the 2 to 12 carbonatoms and at least one carbon-carbon triple bond. Alkynyl groups may beoptionally substituted with one or more substituents.

The sp² or sp carbons of an alkenyl group and an alkynyl group,respectively, may optionally be the point of attachment of the alkenylor alkynyl groups.

The term “alkoxy” refers to an —OR substituent.

As used herein, the term “halogen”, “hal” or “halo” means —F, —Cl, —Bror —I.

The term “haloalkoxy” refers to an —OR substituent where R is fully orpartially substituted with Cl, F, I or Br or any combination thereof.Examples of haloalkoxy groups include trifluoromethoxy, and2,2,2-trifluoroethoxy.

The term “cycloalkyl” refers to a hydrocarbon 3-8 membered monocyclic or7-14 membered bicyclic ring system having at least one saturated ring orhaving at least one non-aromatic ring, wherein the non-aromatic ring mayhave some degree of unsaturation. Cycloalkyl groups may be optionallysubstituted with one or more substituents. In one embodiment, 0, 1, 2,3, or 4 atoms of each ring of a cycloalkyl group may be substituted by asubstituent. Representative examples of cycloalkyl group includecyclopropyl, cyclopentyl, cyclohexyl, cyclobutyl, cycloheptyl,cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, and thelike.

The term “aryl” refers to a hydrocarbon monocyclic, bicyclic ortricyclic aromatic ring system. Aryl groups may be optionallysubstituted with one or more substituents. In one embodiment, 0, 1, 2,3, 4, 5 or 6 atoms of each ring of an aryl group may be substituted by asubstituent. Examples of aryl groups include phenyl, naphthyl,anthracenyl, fluorenyl, indenyl, azulenyl, and the like.

The term “heteroaryl” refers to an aromatic 5-8 membered monocyclic,8-12 membered bicyclic, or 11-14 membered tricyclic ring system having1-4 ring heteroatoms if monocyclic, 1-6 heteroatoms if bicyclic, or 1-9heteroatoms if tricyclic, said heteroatoms selected from O, N, or S, andthe remaining ring atoms being carbon (with appropriate hydrogen atomsunless otherwise indicated). Heteroaryl groups may be optionallysubstituted with one or more substituents. In one embodiment, 0, 1, 2,3, or 4 atoms of each ring of a heteroaryl group may be substituted by asubstituent. Examples of heteroaryl groups include pyridyl, furanyl,thienyl, pyrrolyl, oxazolyl, oxadiazolyl, imidazolyl, thiazolyl,isoxazolyl, quinolinyl, pyrazolyl, isothiazolyl, pyridazinyl,pyrimidinyl, pyrazinyl, triazinyl, isoquinolinyl, indazolyl, and thelike.

The term “nitrogen-containing heteroaryl” refers to a heteroaryl grouphaving 1-4 ring nitrogen heteroatoms if monocyclic, 1-6 ring nitrogenheteroatoms if bicyclic, or 1-9 ring nitrogen heteroatoms if tricyclic.

The term “heterocycloalkyl” refers to a nonaromatic 3-8 memberedmonocyclic, 7-12 membered bicyclic, or 10-14 membered tricyclic ringsystem comprising 1-3 heteroatoms if monocyclic, 1-6 heteroatoms ifbicyclic, or 1-9 heteroatoms if tricyclic, said heteroatoms selectedfrom O, N, S, B, P or Si, wherein the nonaromatic ring system iscompletely saturated. Heterocycloalkyl groups may be optionallysubstituted with one or more substituents. In one embodiment, 0, 1, 2,3, or 4 atoms of each ring of a heterocycloalkyl group may besubstituted by a substituent. Representative heterocycloalkyl groupsinclude piperidinyl, piperazinyl, tetrahydropyranyl, morpholinyl,thiomorpholinyl, 1,3-dioxolane, tetrahydrofuranyl, tetrahydrothienyl,thiirenyl, and the like.

The term “alkylamino” refers to an amino substituent which is furthersubstituted with one or two alkyl groups. The term “aminoalkyl” refersto an alkyl substituent which is further substituted with one or moreamino groups. The term “hydroxyalkyl” or “hydroxylalkyl” refers to analkyl substituent which is further substituted with one or more hydroxylgroups. The alkyl or aryl portion of alkylamino, aminoalkyl,mercaptoalkyl, hydroxyalkyl, mercaptoalkoxy, sulfonylalkyl,sulfonylaryl, alkylcarbonyl, and alkylcarbonylalkyl may be optionallysubstituted with one or more substituents.

Acids and bases useful in the methods herein are known in the art. Acidcatalysts are any acidic chemical, which can be inorganic (e.g.,hydrochloric, sulfuric, nitric acids, aluminum trichloride) or organic(e.g., camphorsulfonic acid, p-toluenesulfonic acid, acetic acid,ytterbium triflate) in nature. Acids are useful in either catalytic orstoichiometric amounts to facilitate chemical reactions. Bases are anybasic chemical, which can be inorganic (e.g., sodium bicarbonate,potassium hydroxide) or organic (e.g., triethylamine, pyridine) innature. Bases are useful in either catalytic or stoichiometric amountsto facilitate chemical reactions.

Alkylating agents are any reagent that is capable of effecting thealkylation of the functional group at issue (e.g., oxygen atom of analcohol, nitrogen atom of an amino group). Alkylating agents are knownin the art, including in the references cited herein, and include alkylhalides (e.g., methyl iodide, benzyl bromide or chloride), alkylsulfates (e.g., methyl sulfate), or other alkyl group-leaving groupcombinations known in the art. Leaving groups are any stable speciesthat can detach from a molecule during a reaction (e.g., eliminationreaction, substitution reaction) and are known in the art, including inthe references cited herein, and include halides (e.g., I—, Cl—, Br—,F—), hydroxy, alkoxy (e.g., —OMe, —O-t-Bu), acyloxy anions (e.g., —OAc,—OC(O)CF₃), sulfonates (e.g., mesyl, tosyl), acetamides (e.g.,—NHC(O)Me), carbamates (e.g., N(Me)C(O)Ot-Bu), phosphonates (e.g.,—OP(O)(OEt)₂), water or alcohols (protic conditions), and the like.

In certain embodiments, substituents on any group (such as, for example,alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl,cycloalkyl, heterocycloalkyl) can be at any atom of that group, whereinany group that can be substituted (such as, for example, alkyl, alkenyl,alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, cycloalkyl,heterocycloalkyl) can be optionally substituted with one or moresubstituents (which may be the same or different), each replacing ahydrogen atom. Examples of suitable substituents include, but are notlimited to alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,aralkyl, heteroaralkyl, aryl, heteroaryl, halogen, haloalkyl, cyano,nitro, alkoxy, aryloxy, hydroxyl, hydroxylalkyl, oxo (i.e., carbonyl),carboxyl, formyl, alkylcarbonyl, alkylcarbonylalkyl, alkoxycarbonyl,alkylcarbonyloxy, aryloxycarbonyl, heteroaryloxy, heteroaryloxycarbonyl,thio, mercapto, mercaptoalkyl, arylsulfonyl, amino, aminoalkyl,dialkylamino, alkylcarbonylamino, alkylaminocarbonyl,alkoxycarbonylamino, alkylamino, arylamino, diarylamino, alkylcarbonyl,or arylamino-substituted aryl; arylalkylamino, aralkylaminocarbonyl,amido, alkylaminosulfonyl, arylaminosulfonyl, dialkylaminosulfonyl,alkylsulfonylamino, arylsulfonylamino, imino, carbamido, carbamyl,thioureido, thiocyanato, sulfoamido, sulfonylalkyl, sulfonylaryl,mercaptoalkoxy, N-hydroxyamidinyl, or N′-aryl, N″-hydroxyamidinyl.

Compounds of the invention can be made by means known in the art oforganic synthesis. Methods for optimizing reaction conditions, ifnecessary minimizing competing by-products, are known in the art.Reaction optimization and scale-up may advantageously utilize high-speedparallel synthesis equipment and computer-controlled microreactors (e.g.Design And Optimization in Organic Synthesis, 2^(nd) Edition, Carlson R,Ed, 2005; Elsevier Science Ltd; Jahnisch, K. et al., Angew. Chem. Int.Ed. Engl. 2004, 43, 406; and references therein). Additional reactionschemes and protocols may be determined by the skilled artisan by use ofcommercially available structure-searchable database software, forinstance, SciFinder® (Chemical Abstracts Service (CAS®) division of theAmerican Chemical Society) and CrossFire Beilstein® (Elsevier MDL), orby appropriate keyword searching using an internet search engine such asGoogle® or keyword databases such as the US Patent and Trademark Officetext database.

The compounds herein may also contain linkages (e.g., carbon-carbonbonds) wherein bond rotation is restricted about that particularlinkage, e.g. restriction resulting from the presence of a ring ordouble bond. Accordingly, all cis/trans and E/Z isomers are expresslyincluded in the present invention. The compounds herein may also berepresented in multiple tautomeric forms, in such instances, theinvention expressly includes all tautomeric forms of the compoundsdescribed herein, even though only a single tautomeric form may berepresented. All such isomeric forms of such compounds herein areexpressly included in the present invention. All crystal forms andpolymorphs of the compounds described herein are expressly included inthe present invention. Also embodied are extracts and fractionscomprising compounds of the invention. The term isomers is intended toinclude diastereoisomers, enantiomers, regioisomers, structural isomers,rotational isomers, tautomers, and the like. For compounds which containone or more stereogenic centers, e.g., chiral compounds, the methods ofthe invention may be carried out with an enantiomerically enrichedcompound, a racemate, or a mixture of diastereomers.

Preferred enantiomerically enriched compounds have an enantiomericexcess of 50% or more, more preferably the compound has an enantiomericexcess of 60%, 70%, 80%, 90%, 95%, 98%, or 99% or more. In preferredembodiments, only one enantiomer or diastereomer of a chiral compound ofthe invention is administered to cells or a subject.

In another aspect, the invention provides a method of synthesizing acompound of formula I (or any of the formulae herein) as describedherein. Another embodiment is a method of making a compound of any ofthe formulae herein using any one, or combination of, reactionsdelineated herein. The method can include the use of one or moreintermediates or chemical reagents delineated herein.

Methods of Treatment

In one aspect, the invention provides a method of modulating themetalloenzyme activity of a cell in a subject, comprising contacting thesubject with a compound of any of the formulae herein (e.g., Formula I),in an amount and under conditions sufficient to modulate metalloenzymeactivity.

In one embodiment, the modulation is inhibition.

In another aspect, the invention provides a method of treating a subjectsuffering from or susceptible to a metalloenzyme-mediated disorder ordisease, comprising administering to the subject an effective amount ofa compound of any of the formulae herein (e.g., Formula I) orpharmaceutical or agricultural composition thereof.

In other aspects, the invention provides a method of treating a subjectsuffering from or susceptible to a metalloenzyme-mediated disorder ordisease, wherein the subject has been identified as in need of treatmentfor a metalloenzyme-mediated disorder or disease, comprisingadministering to said subject in need thereof, an effective amount of acompound of any of the formulae herein (e.g., Formula I) orpharmaceutical or agricultural composition thereof, such that saidsubject is treated for said disorder.

In certain embodiments, the invention provides a method of treating adisease, disorder or symptom thereof, wherein the disorder is cancer,cardiovascular disease, inflammatory disease or infectious disease. Inother embodiments the disease, disorder or symptom thereof is metabolicdisease, ophthalmologic disease, central nervous system (CNS) disease,urologic disease, or gastrointestinal disease. In certain embodimentsthe disease is prostate cancer, breast cancer, inflammatory boweldisease, psoriasis, systemic fungal infection, skin structure fungalinfection, mucosal fungal infection, and onychomycosis.

In certain embodiments, the subject is a mammal, preferably a primate orhuman.

In another embodiment, the invention provides a method as describedabove, wherein the effective amount of the compound of any of theformulae herein (e.g., Formula I) is as described above.

In another embodiment, the invention provides a method as describedabove, wherein the compound of any of the formulae herein (e.g., FormulaI) is administered intravenously, intramuscularly, subcutaneously,intracerebroventricularly, orally or topically.

In other embodiments, the invention provides a method as describedabove, wherein the compound of any of the formulae herein (e.g., FormulaI) is administered alone or in combination with one or more othertherapeutics. In a further embodiment, the additional therapeutic agentis an anti-cancer agent, antifungal agent, cardiovascular agent,antiinflammatory agent, chemotherapeutic agent, an anti-angiogenesisagent, cytotoxic agent, an anti-proliferation agent, metabolic diseaseagent, opthalmologic disease agent, central nervous system (CNS) diseaseagent, urologic disease agent, or gastrointestinal disease agent.

Another object of the present invention is the use of a compound asdescribed herein (e.g., of any formulae herein) in the manufacture of amedicament for use in the treatment of a metalloenzyme-mediated disorderor disease. Another object of the present invention is the use of acompound as described herein (e.g., of any formulae herein) for use inthe treatment of a metalloenzyme-mediated disorder or disease. Anotherobject of the present invention is the use of a compound as describedherein (e.g., of any formulae herein) in the manufacture of anagricultural composition for use in the treatment or prevention of ametalloenzyme-mediated disorder or disease in agricultural or agrariansettings.

Pharmaceutical Compositions

In one aspect, the invention provides a pharmaceutical compositioncomprising the compound of any of the formulae herein (e.g., Formula I)and a pharmaceutically acceptable carrier.

In another embodiment, the invention provides a pharmaceuticalcomposition further comprising an additional therapeutic agent. In afurther embodiment, the additional therapeutic agent is an anti-canceragent, antifungal agent, cardiovascular agent, antiinflammatory agent,chemotherapeutic agent, an anti-angiogenesis agent, cytotoxic agent, ananti-proliferation agent, metabolic disease agent, opthalmologic diseaseagent, central nervous system (CNS) disease agent, urologic diseaseagent, or gastrointestinal disease agent.

In one aspect, the invention provides a kit comprising an effectiveamount of a compound of any of the formulae herein (e.g., Formula I), inunit dosage form, together with instructions for administering thecompound to a subject suffering from or susceptible to ametalloenzyme-mediated disease or disorder, including cancer, solidtumor, cardiovascular disease, inflammatory disease, infectious disease.In other embodiments the disease, disorder or symptom thereof ismetabolic disease, ophthalmologic disease, central nervous system (CNS)disease, urologic disease, or gastrointestinal disease.

The term “pharmaceutically acceptable salts” or “pharmaceuticallyacceptable carrier” is meant to include salts of the active compoundswhich are prepared with relatively nontoxic acids or bases, depending onthe particular substituents found on the compounds described herein.When compounds of the present invention contain relatively acidicfunctionalities, base addition salts can be obtained by contacting theneutral form of such compounds with a sufficient amount of the desiredbase, either neat or in a suitable inert solvent. Examples ofpharmaceutically acceptable base addition salts include sodium,potassium, calcium, ammonium, organic amino, or magnesium salt, or asimilar salt. When compounds of the present invention contain relativelybasic functionalities, acid addition salts can be obtained by contactingthe neutral form of such compounds with a sufficient amount of thedesired acid, either neat or in a suitable inert solvent. Examples ofpharmaceutically acceptable acid addition salts include those derivedfrom inorganic acids like hydrochloric, hydrobromic, nitric, carbonic,monohydrogencarbonic, phosphoric, monohydrogenphosphoric,dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydroiodic, orphosphorous acids and the like, as well as the salts derived fromrelatively nontoxic organic acids like acetic, propionic, isobutyric,maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic,phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric,methanesulfonic, and the like. Also included are salts of amino acidssuch as arginate and the like, and salts of organic acids likeglucuronic or galactunoric acids and the like (see, e.g., Berge et al.,J. Pharm. Sci. 1977, 66, 1-19). Certain specific compounds of thepresent invention contain both basic and acidic functionalities thatallow the compounds to be converted into either base or acid additionsalts. Other pharmaceutically acceptable carriers known to those ofskill in the art are suitable for the present invention.

The neutral forms of the compounds may be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents, but otherwise the salts are equivalent to the parentform of the compound for the purposes of the present invention.

In addition to salt forms, the present invention provides compoundswhich are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentinvention. Additionally, prodrugs can be converted to the compounds ofthe present invention by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the present invention when placed in a transdermal patchreservoir with a suitable enzyme or chemical reagent.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are intended to beencompassed within the scope of the present invention. Certain compoundsof the present invention may exist in multiple crystalline or amorphousforms. In general, all physical forms are equivalent for the usescontemplated by the present invention and are intended to be within thescope of the present invention.

The invention also provides a pharmaceutical composition, comprising aneffective amount of a compound described herein and a pharmaceuticallyacceptable carrier. In an embodiment, compound is administered to thesubject using a pharmaceutically-acceptable formulation, e.g., apharmaceutically-acceptable formulation that provides sustained deliveryof the compound to a subject for at least 12 hours, 24 hours, 36 hours,48 hours, one week, two weeks, three weeks, or four weeks after thepharmaceutically-acceptable formulation is administered to the subject.

Actual dosage levels and time course of administration of the activeingredients in the pharmaceutical compositions of this invention may bevaried so as to obtain an amount of the active ingredient which iseffective to achieve the desired therapeutic response for a particularpatient, composition, and mode of administration, without being toxic(or unacceptably toxic) to the patient.

In use, at least one compound according to the present invention isadministered in a pharmaceutically effective amount to a subject in needthereof in a pharmaceutical carrier by intravenous, intramuscular,subcutaneous, or intracerebroventricular injection or by oraladministration or topical application. In accordance with the presentinvention, a compound of the invention may be administered alone or inconjunction with a second, different therapeutic. By “in conjunctionwith” is meant together, substantially simultaneously or sequentially.In one embodiment, a compound of the invention is administered acutely.The compound of the invention may therefore be administered for a shortcourse of treatment, such as for about 1 day to about 1 week. In anotherembodiment, the compound of the invention may be administered over alonger period of time to ameliorate chronic disorders, such as, forexample, for about one week to several months depending upon thecondition to be treated.

By “pharmaceutically effective amount” as used herein is meant an amountof a compound of the invention, high enough to significantly positivelymodify the condition to be treated but low enough to avoid serious sideeffects (at a reasonable benefit/risk ratio), within the scope of soundmedical judgment. A pharmaceutically effective amount of a compound ofthe invention will vary with the particular goal to be achieved, the ageand physical condition of the patient being treated, the severity of theunderlying disease, the duration of treatment, the nature of concurrenttherapy and the specific compound employed. For example, atherapeutically effective amount of a compound of the inventionadministered to a child or a neonate will be reduced proportionately inaccordance with sound medical judgment. The effective amount of acompound of the invention will thus be the minimum amount which willprovide the desired effect.

A decided practical advantage of the present invention is that thecompound may be administered in a convenient manner such as byintravenous, intramuscular, subcutaneous, oral orintracerebroventricular injection routes or by topical application, suchas in creams or gels. Depending on the route of administration, theactive ingredients which comprise a compound of the invention may berequired to be coated in a material to protect the compound from theaction of enzymes, acids and other natural conditions which mayinactivate the compound. In order to administer a compound of theinvention by other than parenteral administration, the compound can becoated by, or administered with, a material to prevent inactivation.

The compound may be administered parenterally or intraperitoneally.Dispersions can also be prepared, for example, in glycerol, liquidpolyethylene glycols, and mixtures thereof, and in oils.

Some examples of substances which can serve as pharmaceutical carriersare sugars, such as lactose, glucose and sucrose; starches such as cornstarch and potato starch; cellulose and its derivatives such as sodiumcarboxymethylcellulose, ethylcellulose and cellulose acetates; powderedtragancanth; malt; gelatin; talc; stearic acids; magnesium stearate;calcium sulfate; vegetable oils, such as peanut oils, cottonseed oil,sesame oil, olive oil, corn oil and oil of theobroma; polyols such aspropylene glycol, glycerine, sorbitol, mannitol, and polyethyleneglycol; agar; alginic acids; pyrogen-free water; isotonic saline; andphosphate buffer solution; skim milk powder; as well as other non-toxiccompatible substances used in pharmaceutical formulations such asVitamin C, estrogen and Echinacea, for example. Wetting agents andlubricants such as sodium lauryl sulfate, as well as coloring agents,flavoring agents, lubricants, excipients, tableting agents, stabilizers,anti-oxidants and preservatives, can also be present. Solubilizingagents, including for example, cremaphore and beta-cyclodextrins canalso used in the pharmaceutical compositions herein.

Pharmaceutical compositions comprising the active compounds of thepresently disclosed subject matter (or prodrugs thereof) can bemanufactured by means of conventional mixing, dissolving, granulating,dragee-making, levigating, emulsifying, encapsulating, entrapping orlyophilization processes. The compositions can be formulated in aconventional manner using one or more physiologically acceptablecarriers, diluents, excipients or auxiliaries which facilitateprocessing of the active compounds into preparations which can be usedpharmaceutically.

Pharmaceutical compositions of the presently disclosed subject mattercan take a form suitable for virtually any mode of administration,including, for example, topical, ocular, oral, buccal, systemic, nasal,injection, transdermal, rectal, vaginal, and the like, or a formsuitable for administration by inhalation or insufflation.

For topical administration, the active compound(s) or prodrug(s) can beformulated as solutions, gels, ointments, creams, suspensions, and thelike.

Systemic formulations include those designed for administration byinjection, e.g., subcutaneous, intravenous, intramuscular, intrathecalor intraperitoneal injection, as well as those designed for transdermal,transmucosal, oral, or pulmonary administration.

Useful injectable preparations include sterile suspensions, solutions oremulsions of the active compound(s) in aqueous or oily vehicles. Thecompositions also can contain formulating agents, such as suspending,stabilizing and/or dispersing agent. The formulations for injection canbe presented in unit dosage form (e.g., in ampules or in multidosecontainers) and can contain added preservatives.

Alternatively, the injectable formulation can be provided in powder formfor reconstitution with a suitable vehicle, including but not limited tosterile pyrogen-free water, buffer, dextrose solution, and the like,before use. To this end, the active compound(s) can be dried by anyart-known technique, such as lyophilization, and reconstituted prior touse.

For transmucosal administration, penetrants appropriate to the barrierto be permeated are used in the formulation. Such penetrants are knownin the art.

For oral administration, the pharmaceutical compositions can take theform of, for example, lozenges, tablets or capsules prepared byconventional means with pharmaceutically acceptable excipients such asbinding agents (e.g., pregelatinized maize starch, polyvinylpyrrolidoneor hydroxypropyl methylcellulose); fillers (e.g., lactose,microcrystalline cellulose or calcium hydrogen phosphate); lubricants(e.g., magnesium stearate, talc or silica); disintegrants (e.g., potatostarch or sodium starch glycolate); or wetting agents (e.g., sodiumlauryl sulfate). The tablets can be coated by methods well known in theart with, for example, sugars or enteric coatings.

Liquid preparations for oral administration can take the form of, forexample, elixirs, solutions, syrups or suspensions, or they can bepresented as a dry product for constitution with water or other suitablevehicle before use. Such liquid preparations can be prepared byconventional means with pharmaceutically acceptable additives such assuspending agents (e.g., sorbitol syrup, cellulose derivatives orhydrogenated edible fats); emulsifying agents (e.g., lecithin oracacia); non-aqueous vehicles (e.g., almond oil, oily esters, ethylalcohol or fractionated vegetable oils); and preservatives (e.g., methylor propyl p-hydroxybenzoates or sorbic acid). The preparations also cancontain buffer salts, preservatives, flavoring, coloring and sweeteningagents as appropriate.

Preparations for oral administration can be suitably formulated to givecontrolled release of the active compound or prodrug, as is well known.

For buccal administration, the compositions can take the form of tabletsor lozenges formulated in a conventional manner.

For rectal and vaginal routes of administration, the active compound(s)can be formulated as solutions (for retention enemas), suppositories, orointments containing conventional suppository bases, such as cocoabutter or other glycerides.

For nasal administration or administration by inhalation orinsufflation, the active compound(s) or prodrug(s) can be convenientlydelivered in the form of an aerosol spray from pressurized packs or anebulizer with the use of a suitable propellant, e.g.,dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, fluorocarbons, carbon dioxide or othersuitable gas. In the case of a pressurized aerosol, the dosage unit canbe determined by providing a valve to deliver a metered amount. Capsulesand cartridges for use in an inhaler or insufflator (for examplecapsules and cartridges comprised of gelatin) can be formulatedcontaining a powder mix of the compound and a suitable powder base suchas lactose or starch.

A specific example of an aqueous suspension formulation suitable fornasal administration using commercially-available nasal spray devicesincludes the following ingredients: active compound or prodrug (0.5-20mg/mL); benzalkonium chloride (0.1-0.2 mg/mL); polysorbate 80 (TWEEN®80; 0.5-5 mg/mL); carboxymethylcellulose sodium or microcrystallinecellulose (1-15 mg/mL); phenylethanol (1-4 mg/mL); and dextrose (20-50mg/mL). The pH of the final suspension can be adjusted to range fromabout pH 5 to pH 7, with a pH of about pH 5.5 being typical.

For ocular administration, the active compound(s) or prodrug(s) can beformulated as a solution, emulsion, suspension, and the like, suitablefor administration to the eye. A variety of vehicles suitable foradministering compounds to the eye are known in the art. Specificnon-limiting examples are described in U.S. Pat. Nos. 6,261,547;6,197,934; 6,056,950; 5,800,807; 5,776,445; 5,698,219; 5,521,222;5,403,841; 5,077,033; 4,882,150; and 4,738,851, each of which isincorporated herein by reference in its entirety.

For prolonged delivery, the active compound(s) or prodrug(s) can beformulated as a depot preparation for administration by implantation orintramuscular injection. The active ingredient can be formulated withsuitable polymeric or hydrophobic materials (e.g., as an emulsion in anacceptable oil) or ion exchange resins, or as sparingly solublederivatives, e.g., as a sparingly soluble salt. Alternatively,transdermal delivery systems manufactured as an adhesive disc or patchwhich slowly releases the active compound(s) for percutaneous absorptioncan be used. To this end, permeation enhancers can be used to facilitatetransdermal penetration of the active compound(s). Suitable transdermalpatches are described in for example, U.S. Pat. Nos. 5,407,713;5,352,456; 5,332,213; 5,336,168; 5,290,561; 5,254,346; 5,164,189;5,163,899; 5,088,977; 5,087,240; 5,008,110; and 4,921,475, each of whichis incorporated herein by reference in its entirety.

Alternatively, other pharmaceutical delivery systems can be employed.Liposomes and emulsions are well-known examples of delivery vehiclesthat can be used to deliver active compound(s) or prodrug(s). Certainorganic solvents such as dimethylsulfoxide (DMSO) also can be employed.

The pharmaceutical compositions can, if desired, be presented in a packor dispenser device which can contain one or more unit dosage formscontaining the active compound(s). The pack can, for example, comprisemetal or plastic foil, such as a blister pack. The pack or dispenserdevice can be accompanied by instructions for administration.

The active compound(s) or prodrug(s) of the presently disclosed subjectmatter, or compositions thereof, will generally be used in an amounteffective to achieve the intended result, for example in an amounteffective to treat or prevent the particular disease being treated. Thecompound(s) can be administered therapeutically to achieve therapeuticbenefit or prophylactically to achieve prophylactic benefit. Bytherapeutic benefit is meant eradication or amelioration of theunderlying disorder being treated and/or eradication or amelioration ofone or more of the symptoms associated with the underlying disorder suchthat the patient reports an improvement in feeling or condition,notwithstanding that the patient can still be afflicted with theunderlying disorder. For example, administration of a compound to apatient suffering from an allergy provides therapeutic benefit not onlywhen the underlying allergic response is eradicated or ameliorated, butalso when the patient reports a decrease in the severity or duration ofthe symptoms associated with the allergy following exposure to theallergen. As another example, therapeutic benefit in the context ofasthma includes an improvement in respiration following the onset of anasthmatic attack, or a reduction in the frequency or severity ofasthmatic episodes. Therapeutic benefit also includes halting or slowingthe progression of the disease, regardless of whether improvement isrealized.

For prophylactic administration, the compound can be administered to apatient at risk of developing one of the previously described diseases.A patient at risk of developing a disease can be a patient havingcharacteristics placing the patient in a designated group of at riskpatients, as defined by an appropriate medical professional or group. Apatient at risk may also be a patient that is commonly or routinely in asetting where development of the underlying disease that may be treatedby administration of a metalloenzyme inhibitor according to theinvention could occur. In other words, the at risk patient is one who iscommonly or routinely exposed to the disease or illness causingconditions or may be acutely exposed for a limited time. Alternatively,prophylactic administration can be applied to avoid the onset ofsymptoms in a patient diagnosed with the underlying disorder.

The amount of compound administered will depend upon a variety offactors, including, for example, the particular indication beingtreated, the mode of administration, whether the desired benefit isprophylactic or therapeutic, the severity of the indication beingtreated and the age and weight of the patient, the bioavailability ofthe particular active compound, and the like. Determination of aneffective dosage is well within the capabilities of those skilled in theart.

Effective dosages can be estimated initially from in vitro assays. Forexample, an initial dosage for use in animals can be formulated toachieve a circulating blood or serum concentration of active compoundthat is at or above an IC₅₀ of the particular compound as measured in anin vitro assay, such as the in vitro fungal MIC or minimal fungicidalconcentration (MFC) and other in vitro assays described in the Examplessection. Calculating dosages to achieve such circulating blood or serumconcentrations taking into account the bioavailability of the particularcompound is well within the capabilities of skilled artisans. Forguidance, see Fingl & Woodbury, “General Principles,” In: Goodman andGilman's The Pharmaceutical Basis of Therapeutics, Chapter 1, pp. 1-46,12th edition, McGraw-Hill Professional, and the references citedtherein, which are incorporated herein by reference.

Initial dosages also can be estimated from in vivo data, such as animalmodels. Animal models useful for testing the efficacy of compounds totreat or prevent the various diseases described above are well-known inthe art.

Dosage amounts will typically be in the range of from about 0.0001 or0.001 or 0.01 mg/kg/day to about 100 mg/kg/day, but can be higher orlower, depending upon, among other factors, the activity of thecompound, its bioavailability, the mode of administration, and variousfactors discussed above. Dosage amount and interval can be adjustedindividually to provide plasma levels of the compound(s) which aresufficient to maintain therapeutic or prophylactic effect. In cases oflocal administration or selective uptake, such as local topicaladministration, the effective local concentration of active compound(s)cannot be related to plasma concentration. Skilled artisans will be ableto optimize effective local dosages without undue experimentation.

The compound(s) can be administered once per day, a few or several timesper day, or even multiple times per day, depending upon, among otherthings, the indication being treated and the judgment of the prescribingphysician.

Preferably, the compound(s) will provide therapeutic or prophylacticbenefit without causing substantial toxicity. Toxicity of thecompound(s) can be determined using standard pharmaceutical procedures.The dose ratio between toxic and therapeutic (or prophylactic) effect isthe therapeutic index. Compounds(s) that exhibit high therapeuticindices are preferred.

The recitation of a listing of chemical groups in any definition of avariable herein includes definitions of that variable as any singlegroup or combination of listed groups. The recitation of an embodimentfor a variable herein includes that embodiment as any single embodimentor in combination with any other embodiments or portions thereof. Therecitation of an embodiment herein includes that embodiment as anysingle embodiment or in combination with any other embodiments orportions thereof.

Agricultural Applications

Compounds of Formula I may be formulated into agriculturally acceptableacid addition salts. By way of a non-limiting example, an amine functioncan form salts with hydrochloric, hydrobromic, sulfuric, phosphoric,acetic, benzoic, citric, malonic, salicylic, malic, fumaric, oxalic,succinic, tartaric, lactic, gluconic, ascorbic, maleic, aspartic,benzenesulfonic, methanesulfonic, ethanesulfonic,hydroxymethanesulfonic, and hydroxyethanesulfonic acids. Additionally,by way of a non-limiting example, an acid function can form saltsincluding those derived from alkali or alkaline earth metals and thosederived from ammonia and amines. Examples of preferred cations includesodium, potassium, and magnesium.

Compounds of Formula I may be formulated into salt derivatives. By wayof a non-limiting example, a salt derivative can be prepared bycontacting a free base with a sufficient amount of the desired acid toproduce a salt. A free base may be regenerated by treating the salt witha suitable dilute aqueous base solution such as dilute aqueous sodiumhydroxide (NaOH), potassium carbonate, ammonia, and sodium bicarbonate.As an example, in many cases, a pesticide, such as 2,4-D, is made morewater-soluble by converting it to its dimethylamine salt.

Suitable salts include those derived from alkali or alkaline earthmetals and those derived from ammonia and amines. Preferred cationsinclude sodium, potassium, magnesium, and aminium cations of theformula:R¹⁰R¹¹R¹²R¹³N⁺wherein R¹⁰, R¹¹, R¹² and R¹³ each, independently represents hydrogen orC₁-C₁₂ alkyl, C₃-C₁₂ alkenyl or C₃-C₁₂ alkynyl, each of which isoptionally substituted by one or more hydroxy, C₁-C₄ alkoxy, C₁-C₄alkylthio or phenyl groups, provided that R¹⁰, R¹¹, R¹² and R¹³ aresterically compatible. Additionally, any two of R¹⁰, R¹¹, R¹² and R¹³together may represent an aliphatic difunctional moiety containing oneto twelve carbon atoms and up to two oxygen or sulfur atoms. Salts ofthe compounds of Formula I can be prepared by treatment of compounds ofFormula I with a metal hydroxide, such as sodium hydroxide, with anamine, such as ammonia, trimethylamine, diethanolamine,2-methylthiopropylamine, bisallylamine, 2-butoxyethylamine, morpholine,cyclododecylamine, or benzylamine or with a tetraalkylammoniumhydroxide, such as tetramethylammonium hydroxide or choline hydroxideAmine salts are often preferred forms of the compounds of Formula Ibecause they are water-soluble and lend themselves to the preparation ofdesirable aqueous based herbicidal compositions.

The compounds and compositions herein can be used in methods ofmodulating metalloenzyme activity in a microorganism on a plantcomprising contacting a compound (or composition) herein with the plant(e.g., seed, seedling, grass, weed, grain). The compounds andcompositions herein can be used to treat a plant, field or otheragricultural area (e.g., as herbicides, pesticides, growth regulators,etc.) by administering the compound or composition (e.g., contacting,applying, spraying, atomizing, dusting, etc.) to the subject plant,field or other agricultural area. The administration can be either pre-or post-emergence. The administration can be either as a treatment orpreventative regimen.

One aspect is a method of treating or preventing a fungal disease ordisorder in or on a plant comprising contacting a compound (orcomposition) of any of the formulae herein with the plant. Anotheraspect is a method of treating or preventing fungi growth in or on aplant comprising contacting a compound (or composition) of any of theformulae herein with the plant. Another aspect is a method of inhibitingmicroorganisms in or on a plant comprising contacting a compound (orcomposition) of any of the formulae herein with the plant.

The compounds and compositions herein may be used in methods ofpreventing or controlling pathogen induced diseases on a plantcomprising contacting a compound herein with the plant (e.g., seed,seedling, grass, weed, grain) or an area adjacent to the plant. Thecompounds and compositions herein may be used to treat a plant, field orother agricultural area by administering the compound or composition(e.g., contacting, applying, spraying, atomizing, dusting, etc.) to thesubject plant, field or other agricultural area. The administration maybe either pre- or post-emergence. The administration may be either as atreatment or preventative regimen. As such, the compounds, compositionsand agricultural uses herein include lawn, turf, ornamental vegetation,home and garden, farming, range and pasture applications. The pathogenmay be any on a plant and include those delineated herein.

One embodiment of the present disclosure is a use of a compound ofFormula I, for protection of a plant against attack by a phytopathogenicorganism or the treatment of a plant infested by a phytopathogenicorganism, comprising the application of a compound of Formula I, or acomposition comprising the compound to soil, a plant, a part of a plant,foliage, and/or seeds.

Additionally, another embodiment of the present disclosure is acomposition useful for protecting a plant against attack by aphytopathogenic organism and/or treatment of a plant infested by aphytopathogenic organism comprising a compound of Formula I and aphytologically acceptable carrier material.

The compounds of the present disclosure may be applied by any of avariety of known techniques, either as the compounds or as formulationscomprising the compounds. For example, the compounds may be applied tothe roots, seeds or foliage of plants for the control of various fungi,without damaging the commercial value of the plants.

The compounds herein can be used alone or in combination with otheragriculturally active agents. The use of the compounds or compositions(and the compositions) herein can further comprise an additional activeagent such as an azole fungicide selected from epoxiconazole,tebuconazole, fluquinconazole, flutriafol, metconazole, myclobutanil,cycproconazole, prothioconazole and propiconazole.

The use of the compounds or compositions (and the compositions) hereincan further comprise an additional active agent such as an azolefungicide selected from the group trifloxystrobin, pyraclostrobin,orysastrobin, fluoxastrobin and azoxystrobin.

Preferably, the compounds of the present disclosure are applied in theform of a formulation, comprising one or more of the compounds ofFormula I with an agriculturally or phytologically acceptable carrier.The compositions comprising compounds herein can be employed, forexample, in the form of directly sprayable aqueous solutions, powders,suspensions, also highly-concentrated aqueous, oily or other suspensionsor dispersions, emulsions, oil dispersions, pastes, dusts, materials forspreading or granules, by means of spraying, atomizing, dusting,spreading or pouring.

The present disclosure contemplates all vehicles by which one or more ofthe compounds may be formulated for delivery and use as a fungicide.Typically, formulations are applied as aqueous suspensions or emulsions.Aqueous use forms can be prepared from emulsion concentrates,suspensions, pastes, wettable powders or water-dispersible granules byadding water. To prepare emulsions, pastes or oil dispersions, thesubstances, as such or dissolved in an oil or solvent, can behomogenized in water by means of wetting agent, tackifier, dispersant oremulsifier. However, it is also possible to prepare concentratescomposed of active substance, wetting agent, tackifier, dispersant oremulsifier and, if appropriate, solvent or oil, and these concentratesare suitable for dilution with water.

Wettable powders, which may be compacted to form water dispersiblegranules, comprise an intimate mixture of one or more of the compoundsof Formula I, an inert carrier and surfactants. The concentration of thecompound in the wettable powder may be from about 10 percent to about 90percent by weight based on the total weight of the wettable powder, morepreferably about 25 weight percent to about 75 weight percent. In thepreparation of wettable powder formulations, the compounds may becompounded with any finely divided solid, such as prophyllite, talc,chalk, gypsum, Fuller's earth, bentonite, attapulgite, starch, casein,gluten, montmorillonite clays, diatomaceous earths, purified silicatesor the like. In such operations, the finely divided carrier andsurfactants are typically blended with the compound(s) and milled.

Granules, e.g. coated granules, impregnated granules and homogeneousgranules, can be prepared by binding the active ingredients (e.g.,compounds herein) to solid carriers. Solid carriers are mineral earthssuch as silicas, silica gels, silicates, talc, kaolin, limestone, lime,chalk, bole, loess, clay, dolomite, diatomaceous earth, calcium sulfate,magnesium sulfate, magnesium oxide, ground synthetic material,fertilizers such as ammonium sulfate, ammonium phosphate, ammoniumnitrate, ureas and products of vegetable origin such as cereal meal,tree bark meal, wood meal and nutshell meal, cellulose powders or othersolid carriers.

The compounds herein can be formulated as ordinary tablets, capsules,solids, liquids, emulsions, slurries, oils, fine granules or powders,which are suitable for administration to plants, fields or otheragricultural areas. In preferred embodiments, the preparation includesbetween 1 and 95% (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 25%, 75%, 80%,90%, 95%) compound herein in a carrier or diluent. The compositionsdelineated herein include the compounds of the formulae delineatedherein, as well as additional agricultural agents if present, in amountseffective for controlling (e.g., modulating, inhibiting) ametalloenzyme-mediated agricultural disease or disorder.

In one approach, a compound herein is provided in an encapsulatedformulation (liquid or powder). Specific materials suitable for use incapsule materials include, but are not limited to, porous particulatesor substrates such as silica, perlite, talc, clay, pyrophyllite,diatomaceous earth, gelatin and gels, polymers (e.g., polyurea,polyurethane, polyamide, polyester, etc.), polymeric particles, orcellulose. These include, for example, hollow fibers, hollow tubes ortubing which release a compound specified herein through the walls,capillary tubing which releases the compound out of an opening in thetubing, polymeric blocks of different shapes, e.g., strips, blocks,tablets, discs, which release the compound out of the polymer matrix,membrane systems which hold the compound within an impermeable containerand release it through a measured permeable membrane, and combinationsof the foregoing. Examples of such dispensing compositions are polymerlaminates, polyvinyl chloride pellets, and microcapillaries.

Encapsulation processes are typically classified as chemical ormechanical. Examples of chemical processes for encapsulation include,but are not limited to, complex coacervation, polymer-polymerincompatibility, interfacial polymerization in liquid media, in situpolymerization, in-liquid drying, thermal and ionic gelation in liquidmedia, desolvation in liquid media, starch-based chemistry processes,trapping in cyclodextrins, and formation of liposomes. Examples ofmechanical processes for encapsulation include, but are not limited to,spray drying, spray chilling, fluidized bed, electrostatic deposition,centrifugal extrusion, spinning disk or rotational suspensionseparation, annular-jet encapsulation, polymerization at liquid-gas orsolid-gas interface, solvent evaporation, pressure extrusion or sprayinginto solvent extraction bath.

Microcapsules are also suitable for the long-term release of activecompound herein. Microcapsules are small particles that contain a corematerial or active ingredient surrounded by a coating or shell. The sizeof the microcapsule typically varies from 1 to 1000 microns withcapsules smaller than 1 micron classified as nanocapsules and capsuleslarger than 1000 microns as macrocapsules. Core payload usually variesfrom 0.1 to 98 weight percent. Microcapsules can have a variety ofstructures (continuous core/shell, multinuclear, or monolithic) and haveirregular or geometric shapes.

In another approach, the compound herein is provided in an oil-baseddelivery system. Oil release substrates include vegetable and/or mineraloils. In one embodiment, the substrate also contains a surface activeagent that renders the composition readily dispersable in water; suchagents include wetting agents, emulsifying agents, dispersing agents,and the like.

Compounds of the invention can also be provided as emulsions. Emulsionformulations can be found as water in oil (w/o) or oil in water (o/w).Droplet size can vary from the nanometer scale (colloidal dispersion) toseveral hundred microns. A variety of surfactants and thickeners areusually incorporated in the formulation to modify the size of thedroplets, stabilize the emulsion, and modify the release.

Emulsifiable concentrates of the compounds of Formula I may comprise aconvenient concentration, such as from about 10 weight percent to about50 weight percent of the compound, in a suitable liquid, based on thetotal weight of the concentrate. The compounds may be dissolved in aninert carrier, which is either a water-miscible solvent or a mixture ofwater-immiscible organic solvents, and emulsifiers. The concentrates maybe diluted with water and oil to form spray mixtures in the form ofoil-in-water emulsions. Useful organic solvents include aromatics,especially the high-boiling naphthalenic and olefinic portions ofpetroleum such as heavy aromatic naphtha. Other organic solvents mayalso be used, for example, terpenic solvents, including rosinderivatives, aliphatic ketones, such as cyclohexanone, and complexalcohols, such as 2-ethoxyethanol.

Emulsifiers which may be advantageously employed herein may be readilydetermined by those skilled in the art and include various nonionic,anionic, cationic and amphoteric emulsifiers, or a blend of two or moreemulsifiers. Examples of nonionic emulsifiers useful in preparing theemulsifiable concentrates include the polyalkylene glycol ethers andcondensation products of alkyl and aryl phenols, aliphatic alcohols,aliphatic amines or fatty acids with ethylene oxide, propylene oxidessuch as the ethoxylated alkyl phenols and carboxylic esters solubilizedwith the polyol or polyoxyalkylene. Cationic emulsifiers includequaternary ammonium compounds and fatty amine salts. Anionic emulsifiersinclude the oil-soluble salts (e.g., calcium) of alkylaryl sulfonicacids, oil-soluble salts or sulfated polyglycol ethers and appropriatesalts of phosphated polyglycol ether.

Representative organic liquids which may be employed in preparing theemulsifiable concentrates of the compounds of the present invention arethe aromatic liquids such as xylene, propyl benzene fractions; or mixednaphthalene fractions, mineral oils, substituted aromatic organicliquids such as dioctyl phthalate; kerosene; dialkyl amides of variousfatty acids, particularly the dimethyl amides of fatty glycols andglycol derivatives such as the n-butyl ether, ethyl ether or methylether of diethylene glycol, the methyl ether of triethylene glycol,petroleum fractions or hydrocarbons such as mineral oil, aromaticsolvents, paraffinic oils, and the like; vegetable oils such as soybeanoil, rapeseed oil, olive oil, castor oil, sunflower seed oil, coconutoil, corn oil, cottonseed oil, linseed oil, palm oil, peanut oil,safflower oil, sesame oil, tung oil and the like; esters of the abovevegetable oils; and the like. Mixtures of two or more organic liquidsmay also be employed in the preparation of the emulsifiable concentrate.Organic liquids include xylene, and propyl benzene fractions, withxylene being most preferred in some cases. Surface-active dispersingagents are typically employed in liquid formulations and in an amount offrom 0.1 to 20 percent by weight based on the combined weight of thedispersing agent with one or more of the compounds. The formulations canalso contain other compatible additives, for example, plant growthregulators and other biologically active compounds used in agriculture.

Aqueous suspensions comprise suspensions of one or more water-insolublecompounds of Formula I, dispersed in an aqueous vehicle at aconcentration in the range from about 5 to about 50 weight percent,based on the total weight of the aqueous suspension. Suspensions areprepared by finely grinding one or more of the compounds, and vigorouslymixing the ground material into a vehicle comprised of water andsurfactants chosen from the same types discussed above. Othercomponents, such as inorganic salts and synthetic or natural gums, mayalso be added to increase the density and viscosity of the aqueousvehicle. It is often most effective to grind and mix at the same time bypreparing the aqueous mixture and homogenizing it in an implement suchas a sand mill, ball mill, or piston-type homogenizer.

Aqueous emulsions comprise emulsions of one or more water-insolublepesticidally active ingredients emulsified in an aqueous vehicle at aconcentration typically in the range from about 5 to about 50 weightpercent, based on the total weight of the aqueous emulsion. If thepesticidally active ingredient is a solid, it must be dissolved in asuitable water-immiscible solvent prior to the preparation of theaqueous emulsion. Emulsions are prepared by emulsifying the liquidpesticidally active ingredient or water-immiscible solution thereof intoan aqueous medium typically with inclusion of surfactants that aid inthe formation and stabilization of the emulsion as described above. Thisis often accomplished with the aid of vigorous mixing provided by highshear mixers or homogenizers.

The compounds of Formula I can also be applied as granular formulations,which are particularly useful for applications to the soil. Granularformulations generally contain from about 0.5 to about 10 weightpercent, based on the total weight of the granular formulation of thecompound(s), dispersed in an inert carrier which consists entirely or inlarge part of coarsely divided inert material such as attapulgite,bentonite, diatomite, clay or a similar inexpensive substance. Suchformulations are usually prepared by dissolving the compounds in asuitable solvent and applying it to a granular carrier which has beenpreformed to the appropriate particle size, in the range of from about0.5 to about 3 mm A suitable solvent is a solvent in which the compoundis substantially or completely soluble. Such formulations may also beprepared by making a dough or paste of the carrier and the compound andsolvent, and crushing and drying to obtain the desired granularparticle.

Alternatively, compounds of the invention may also be formulated in asolid tablet and comprise (and preferably consist essentially of) anoil, a protein/carbohydrate material (preferably vegetable based), asweetener and an active ingredient useful in the prevention or treatmentof a metalloenzyme-mediated agricultural disease or disorder. In oneembodiment the invention provides a solid tablet and comprises (andpreferably consist essentially of) an oil, a protein/carbohydratematerial (preferably vegetable based), a sweetener and an activeingredient (e.g., compound herein or combinations or derivativesthereof) useful in the prevention or treatment a metalloenzyme-mediatedagricultural disease or disorder. Tablets typically contain about 4-40%(e.g., 5%, 10%, 20%, 30%, 40%) by weight of an oil (e.g., plant oil,such as corn, sunflower, peanut, olive, grape seed, tung, turnip,soybean, cottonseed, walnut, palm, castor, earth almond, hazelnut,avocado, sesame, croton tiglium, cacao, linseed, rapeseed, and canolaoils and their hydrogenated derivatives; petroleum derived oils (e.g.,paraffins and petroleum jelly), and other water immiscible hydrocarbons(e.g., paraffins). The tablets further contain from about 5-40% (e.g.,5%, 10%, 20%, 30%, 40%) by weight of a vegetable-basedprotein/carbohydrate material. The material contains both a carbohydrateportion (e.g., derived from cereal grains, such as wheat, rye, barley,oat, corn, rice, millet, sorghum, birdseed, buckwheat, alfalfa, mielga,corn meal, soybean meal, grain flour, wheat middlings, wheat bran, corngluten meal, algae meal, dried yeast, beans, rice) and a proteinportion.

Optionally, various excipients and binders can be used in order toassist with delivery of the active ingredient or to provide theappropriate structure to the tablet. Preferred excipients and bindersinclude anhydrous lactose, microcrystalline cellulose, corn starch,magnesium estearate, calcium estearate, zinc estearate, sodiumcarboxymethylcellulose, ethyl cellulose, hydroxypropyl methyl cellulose,and mixtures thereof.

Dusts containing the compounds of Formula I may be prepared byintimately mixing one or more of the compounds in powdered form with asuitable dusty agricultural carrier, such as, for example, kaolin clay,ground volcanic rock, and the like. Dusts can suitably contain fromabout 1 to about 10 weight percent of the compounds, based on the totalweight of the dust.

The formulations may additionally contain adjuvant surfactants toenhance deposition, wetting and penetration of the compounds onto thetarget crop and organism. These adjuvant surfactants may optionally beemployed as a component of the formulation or as a tank mix. The amountof adjuvant surfactant will typically vary from 0.01 to 1.0 percent byvolume, based on a spray-volume of water, preferably 0.05 to 0.5 volumepercent. Suitable adjuvant surfactants include, but are not limited toethoxylated nonyl phenols, ethoxylated synthetic or natural alcohols,salts of the esters or sulfosuccinic acids, ethoxylated organosilicones,ethoxylated fatty amines, blends of surfactants with mineral orvegetable oils, crop oil concentrate (mineral oil (85%)+emulsifiers(15%)); nonylphenol ethoxylate; benzylcocoalkyldimethyl quaternaryammonium salt; blend of petroleum hydrocarbon, alkyl esters, organicacid, and anionic surfactant; C₉-C₁₁ alkylpolyglycoside; phosphatedalcohol ethoxylate; natural primary alcohol (C₁₂-C₁₆) ethoxylate;di-sec-butylphenol EO-PO block copolymer; polysiloxane-methyl cap;nonylphenol ethoxylate+urea ammonium nitrate; emulsified methylated seedoil; tridecyl alcohol (synthetic) ethoxylate (8EO); tallow amineethoxylate (15 EO); PEG(400) dioleate-99. The formulations may alsoinclude oil-in-water emulsions such as those disclosed in U.S. patentapplication Ser. No. 11/495,228, the disclosure of which is expresslyincorporated by reference herein.

The formulations may optionally include combinations that contain otherpesticidal compounds. Such additional pesticidal compounds may befungicides, insecticides, herbicides, nematocides, miticides,arthropodicides, bactericides or combinations thereof that arecompatible with the compounds of the present invention in the mediumselected for application, and not antagonistic to the activity of thepresent compounds. Accordingly, in such embodiments, the otherpesticidal compound is employed as a supplemental toxicant for the sameor for a different pesticidal use. The compounds of Formula I and thepesticidal compound in the combination can generally be present in aweight ratio of from 1:100 to 100:1.

The compounds of the present disclosure may also be combined with otherfungicides to form fungicidal mixtures and synergistic mixtures thereof.The fungicidal compounds of the present disclosure are often applied inconjunction with one or more other fungicides to control a wider varietyof undesirable diseases. When used in conjunction with otherfungicide(s), the presently claimed compounds may be formulated with theother fungicide(s), tank mixed with the other fungicide(s) or appliedsequentially with the other fungicide(s). Such other fungicides mayinclude 2-(thiocyanatomethylthio)-benzothiazole, 2-phenylphenol,8-hydroxyquinoline sulfate, ametoctradin, amisulbrom, antimycin,Ampelomyces quisqualis, azaconazole, azoxystrobin, Bacillus subtilis,benalaxyl, benomyl, benthiavalicarb-isopropyl,benzylaminobenzene-sulfonate (BABS) salt, bicarbonates, biphenyl,bismerthiazol, bitertanol, bixafen, blasticidin-S, borax, Bordeauxmixture, boscalid, bromuconazole, bupirimate, calcium polysulfide,captafol, captan, carbendazim, carboxin, carpropamid, carvone,chloroneb, chlorothalonil, chlozolinate, Coniothyrium minitans, copperhydroxide, copper octanoate, copper oxychloride, copper sulfate, coppersulfate (tribasic), cuprous oxide, cyazofamid, cyflufenamid, cymoxanil,cyproconazole, cyprodinil, dazomet, debacarb, diammoniumethylenebis-(dithiocarbamate), dichlofluanid, dichlorophen, diclocymet,diclomezine, dichloran, diethofencarb, difenoconazole, difenzoquat ion,diflumetorim, dimethomorph, dimoxystrobin, diniconazole, diniconazole-M,dinobuton, dinocap, diphenylamine, dithianon, dodemorph, dodemorphacetate, dodine, dodine free base, edifenphos, enestrobin,epoxiconazole, ethaboxam, ethoxyquin, etridiazole, famoxadone,fenamidone, fenarimol, fenbuconazole, fenfuram, fenhexamid, fenoxanil,fenpiclonil, fenpropidin, fenpropimorph, fenpyrazamine, fentin, fentinacetate, fentin hydroxide, ferbam, ferimzone, fluazinam, fludioxonil,flumorph, fluopicolide, fluopyram, fluoroimide, fluoxastrobin,fluquinconazole, flusilazole, flusulfamide, flutianil, flutolanil,flutriafol, fluxapyroxad, folpet, formaldehyde, fosetyl,fosetyl-aluminium, fuberidazole, furalaxyl, furametpyr, guazatine,guazatine acetates, GY-81, hexachlorobenzene, hexaconazole, hymexazol,imazalil, imazalil sulfate, imibenconazole, iminoctadine, iminoctadinetriacetate, iminoctadine tris(albesilate), iodocarb, ipconazole,ipfenpyrazolone, iprobenfos, iprodione, iprovalicarb, isoprothiolane,isopyrazam, isotianil, laminarin, kasugamycin, kasugamycin hydrochloridehydrate, kresoxim-methyl, mancopper, mancozeb, mandipropamid, maneb,mefenoxam, mepanipyrim, mepronil, meptyl-dinocap, mercuric chloride,mercuric oxide, mercurous chloride, metalaxyl, metalaxyl-M, metam,metam-ammonium, metam-potassium, metam-sodium, metconazole,methasulfocarb, methyl iodide, methyl isothiocyanate, metiram,metominostrobin, metrafenone, mildiomycin, myclobutanil, nabam,nitrothal-isopropyl, nuarimol, octhilinone, ofurace, oleic acid (fattyacids), orysastrobin, oxadixyl, oxine-copper, oxpoconazole fumarate,oxycarboxin, pefurazoate, penconazole, pencycuron, penflufen,pentachlorophenol, pentachlorophenyl laurate, penthiopyrad,phenylmercury acetate, phosphonic acid, phthalide, picoxystrobin,polyoxin B, polyoxins, polyoxorim, potassium bicarbonate, potassiumhydroxyquinoline sulfate, probenazole, prochloraz, procymidone,propamocarb, propamocarb hydrochloride, propiconazole, propineb,proquinazid, prothioconazole, pyraclostrobin, pyrametostrobin,pyraoxystrobin, pyrazophos, pyribencarb, pyributicarb, pyrifenox,pyrimethanil, pyriofenone, pyroquilon, quinoclamine, quinoxyfen,quintozene, Reynoutria sachalinensis extract, sedaxane, silthiofam,simeconazole, sodium 2-phenylphenoxide, sodium bicarbonate, sodiumpentachlorophenoxide, spiroxamine, sulfur, SYP-Z071, SYP-Z048, tar oils,tebuconazole, tebufloquin, tecnazene, tetraconazole, thiabendazole,thifluzamide, thiophanate-methyl, thiram, tiadinil, tolclofos-methyl,tolylfluanid, triadimefon, triadimenol, triazoxide, tricyclazole,tridemorph, trifloxystrobin, triflumizole, triforine, triticonazole,validamycin, valifenalate, valiphenal, vinclozolin, zineb, ziram,zoxamide, Candida oleophila, Fusarium oxysporum, Gliocladium spp.,Phlebiopsis gigantea, Streptomyces griseoviridis, Trichoderma spp.,(RS)-N-(3,5-dichlorophenyl)-2-(methoxymethyl)-succinimide,1,2-dichloropropane, 1,3-dichloro-1,1,3,3-tetrafluoroacetone hydrate,1-chloro-2,4-dinitronaphthalene, 1-chloro-2-nitropropane,2-(2-heptadecyl-2-imidazolin-1-yl)ethanol,2,3-dihydro-5-phenyl-1,4-dithi-ine 1,1,4,4-tetraoxide,2-methoxyethylmercury acetate, 2-methoxyethylmercury chloride,2-methoxyethylmercury silicate, 3-(4-chlorophenyl)-5-methylrhodanine,4-(2-nitroprop-1-enyl)phenyl thiocyanateme, ampropylfos, anilazine,azithiram, barium polysulfide, Bayer 32394, benodanil, benquinox,bentaluron, benzamacril; benzamacril-isobutyl, benzamorf, binapacryl,bis(methylmercury) sulfate, bis(tributyltin) oxide, buthiobate, cadmiumcalcium copper zinc chromate sulfate, carbamorph, CECA, chlobenthiazone,chloraniformethan, chlorfenazole, chlorquinox, climbazole, cyclafuramid,cypendazole, cyprofuram, decafentin, dichlone, dichlozoline,diclobutrazol, dimethirimol, dinocton, dinosulfon, dinoterbon,dipyrithione, ditalimfos, dodicin, drazoxolon, EBP, ESBP, etaconazole,etem, ethirim, fenaminosulf, fenapanil, fenitropan, fluotrimazole,furcarbanil, furconazole, furconazole-cis, furmecyclox, furophanate,glyodine, griseofulvin, halacrinate, Hercules 3944, hexylthiofos,ICIA0858, isopamphos, isovaledione, mebenil, mecarbinzid, metazoxolon,methfuroxam, methylmercury dicyandiamide, metsulfovax, milneb,mucochloric anhydride, myclozolin, N-3,5-dichlorophenyl-succinimide,N-3-nitrophenylitaconimide, natamycin,N-ethylmercurio-4-toluenesulfonanilide, nickelbis(dimethyldithiocarbamate), OCH, phenylmercurydimethyldithiocarbamate, phenylmercury nitrate, phosdiphen, picolinamideUK-2A and derivatives thereof, prothiocarb; prothiocarb hydrochloride,pyracarbolid, pyridinitril, pyroxychlor, pyroxyfur, quinacetol,quinacetol sulfate, quinazamid, quinconazole, rabenzazole,salicylanilide, SSF-109, sultropen, tecoram, thiadifluor, thicyofen,thiochlorfenphim, thiophanate, thioquinox, tioxymid, triamiphos,triarimol, triazbutil, trichlamide, urbacid, and zarilamide, and anycombinations thereof.

Additionally, the compounds of the present invention may be combinedwith other pesticides, including insecticides, nematocides, miticides,arthropodicides, bactericides or combinations thereof that arecompatible with the compounds of the present invention in the mediumselected for application, and not antagonistic to the activity of thepresent compounds to form pesticidal mixtures and synergistic mixturesthereof. The fungicidal compounds of the present disclosure may beapplied in conjunction with one or more other pesticides to control awider variety of undesirable pests. When used in conjunction with otherpesticides, the presently claimed compounds may be formulated with theother pesticide(s), tank mixed with the other pesticide(s) or appliedsequentially with the other pesticide(s). Typical insecticides include,but are not limited to: 1,2-dichloropropane, abamectin, acephate,acetamiprid, acethion, acetoprole, acrinathrin, acrylonitrile,alanycarb, aldicarb, aldoxycarb, aldrin, allethrin, allosamidin,allyxycarb, alpha-cypermethrin, alpha-ecdysone, alpha-endosulfan,amidithion, aminocarb, amiton, amiton oxalate, amitraz, anabasine,athidathion, azadirachtin, azamethiphos, azinphos-ethyl,azinphos-methyl, azothoate, barium hexafluorosilicate, barthrin,bendiocarb, benfuracarb, bensultap, beta-cyfluthrin, beta-cypermethrin,bifenthrin, bioallethrin, bioethanomethrin, biopermethrin,bistrifluoron, borax, boric acid, bromfenvinfos, bromocyclen, bromo-DDT,bromophos, bromophos-ethyl, bufencarb, buprofezin, butacarb,butathiofos, butocarboxim, butonate, butoxycarboxim, cadusafos, calciumarsenate, calcium polysulfide, camphechlor, carbanolate, carbaryl,carbofuran, carbon disulfide, carbon tetrachloride, carbophenothion,carbosulfan, cartap, cartap hydrochloride, chlorantraniliprole,chlorbicyclen, chlordane, chlordecone, chlordimeform, chlordimeformhydrochloride, chlorethoxyfos, chlorfenapyr, chlorfenvinphos,chlorfluazuron, chlormephos, chloroform, chloropicrin, chlorphoxim,chlorprazophos, chlorpyrifos, chlorpyrifos-methyl, chlorthiophos,chromafenozide, cinerin I, cinerin II, cinerins, cismethrin,cloethocarb, closantel, clothianidin, copper acetoarsenite, copperarsenate, copper naphthenate, copper oleate, coumaphos, coumithoate,crotamiton, crotoxyphos, crufomate, cryolite, cyanofenphos, cyanophos,cyanthoate, cyantraniliprole, cyclethrin, cycloprothrin, cyfluthrin,cyhalothrin, cypermethrin, cyphenothrin, cyromazine, cythioate, DDT,decarbofuran, deltamethrin, demephion, demephion-O, demephion-S,demeton, demeton-methyl, demeton-O, demeton-O-methyl, demeton-S,demeton-S-methyl, demeton-S-methylsulphon, diafenthiuron, dialifos,diatomaceous earth, diazinon, dicapthon, dichlofenthion, dichlorvos,dicresyl, dicrotophos, dicyclanil, dieldrin, diflubenzuron, dilor,dimefluthrin, dimefox, dimetan, dimethoate, dimethrin, dimethylvinphos,dimetilan, dinex, dinex-diclexine, dinoprop, dinosam, dinotefuran,diofenolan, dioxabenzofos, dioxacarb, dioxathion, disulfoton,dithicrofos, d-limonene, DNOC, DNOC-ammonium, DNOC-potassium,DNOC-sodium, doramectin, ecdysterone, emamectin, emamectin benzoate,EMPC, empenthrin, endosulfan, endothion, endrin, EPN, epofenonane,eprinomectin, esdepallethrine, esfenvalerate, etaphos, ethiofencarb,ethion, ethiprole, ethoate-methyl, ethoprophos, ethyl formate,ethyl-DDD, ethylene dibromide, ethylene dichloride, ethylene oxide,etofenprox, etrimfos, EXD, famphur, fenamiphos, fenazaflor,fenchlorphos, fenethacarb, fenfluthrin, fenitrothion, fenobucarb,fenoxacrim, fenoxycarb, fenpirithrin, fenpropathrin, fensulfothion,fenthion, fenthion-ethyl, fenvalerate, fipronil, flometoquin,flonicamid, flubendiamide, flucofuron, flucycloxuron, flucythrinate,flufenerim, flufenoxuron, flufenprox, flufiprole, flupyradifurone,fluvalinate, fonofos, formetanate, formetanate hydrochloride,formothion, formparanate, formparanate hydrochloride, fosmethilan,fospirate, fosthietan, furathiocarb, furethrin, gamma-cyhalothrin,gamma-HCH, halfenprox, halofenozide, HCH, HEOD, heptachlor, heptenophos,heterophos, hexaflumuron, HHDN, hydramethylnon, hydrogen cyanide,hydroprene, hyquincarb, imidacloprid, imiprothrin, indoxacarb,iodomethane, IPSP, isazofos, isobenzan, isocarbophos, isodrin,isofenphos, isofenphos-methyl, isoprocarb, isoprothiolane, isothioate,isoxathion, ivermectin, jasmolin I, jasmolin II, jodfenphos, juvenilehormone I, juvenile hormone II, juvenile hormone III, kelevan,kinoprene, lambda-cyhalothrin, lead arsenate, lepimectin, leptophos,lindane, lirimfos, lufenuron, lythidathion, malathion, malonoben,mazidox, mecarbam, mecarphon, menazon, meperfluthrin, mephosfolan,mercurous chloride, mesulfenfos, metaflumizone, methacrifos,methamidophos, methidathion, methiocarb, methocrotophos, methomyl,methoprene, methoxychlor, methoxyfenozide, methyl bromide, methylisothiocyanate, methylchloroform, methylene chloride, metofluthrin,metolcarb, metoxadiazone, mevinphos, mexacarbate, milbemectin,milbemycin oxime, mipafox, mirex, molosultap, monocrotophos, monomehypo,monosultap, morphothion, moxidectin, naftalofos, naled, naphthalene,nicotine, nifluridide, nitenpyram, nithiazine, nitrilacarb, novaluron,noviflumuron, omethoate, oxamyl, oxydemeton-methyl, oxydeprofos,oxydisulfoton, para-dichlorobenzene, parathion, parathion-methyl,penfluoron, pentachlorophenol, permethrin, phenkapton, phenothrin,phenthoate, phorate, phosalone, phosfolan, phosmet, phosnichlor,phosphamidon, phosphine, phoxim, phoxim-methyl, pirimetaphos,pirimicarb, pirimiphos-ethyl, pirimiphos-methyl, potassium arsenite,potassium thiocyanate, pp′-DDT, prallethrin, precocene I, precocene II,precocene III, primidophos, profenofos, profluralin, promacyl,promecarb, propaphos, propetamphos, propoxur, prothidathion, prothiofos,prothoate, protrifenbute, pyraclofos, pyrafluprole, pyrazophos,pyresmethrin, pyrethrin I, pyrethrin II, pyrethrins, pyridaben,pyridalyl, pyridaphenthion, pyrifluquinazon, pyrimidifen, pyrimitate,pyriprole, pyriproxyfen, quassia, quinalphos, quinalphos-methyl,quinothion, rafoxanide, resmethrin, rotenone, ryania, sabadilla,schradan, selamectin, silafluofen, silica gel, sodium arsenite, sodiumfluoride, sodium hexafluorosilicate, sodium thiocyanate, sophamide,spinetoram, spinosad, spiromesifen, spirotetramat, sulcofuron,sulcofuron-sodium, sulfluramid, sulfotep, sulfoxaflor, sulfurylfluoride, sulprofos, tau-fluvalinate, tazimcarb, TDE, tebufenozide,tebufenpyrad, tebupirimfos, teflubenzuron, tefluthrin, temephos, TEPP,terallethrin, terbufos, tetrachloroethane, tetrachlorvinphos,tetramethrin, tetramethylfluthrin, theta-cypermethrin, thiacloprid,thiamethoxam, thicrofos, thiocarboxime, thiocyclam, thiocyclam oxalate,thiodicarb, thiofanox, thiometon, thiosultap, thiosultap-disodium,thiosultap-monosodium, thuringiensin, tolfenpyrad, tralomethrin,transfluthrin, transpermethrin, triarathene, triazamate, triazophos,trichlorfon, trichlormetaphos-3, trichloronat, trifenofos, triflumuron,trimethacarb, triprene, vamidothion, vaniliprole, XMC, xylylcarb,zeta-cypermethrin, zolaprofos, and any combinations thereof.

Additionally, the compounds of the present invention may be combinedwith herbicides that are compatible with the compounds of the presentinvention in the medium selected for application, and not antagonisticto the activity of the present compounds to form pesticidal mixtures andsynergistic mixtures thereof. The fungicidal compounds of the presentdisclosure may be applied in conjunction with one or more herbicides tocontrol a wide variety of undesirable plants. When used in conjunctionwith herbicides, the presently claimed compounds may be formulated withthe herbicide(s), tank mixed with the herbicide(s) or appliedsequentially with the herbicide(s). Typical herbicides include, but arenot limited to: 4-CPA; 4-CPB; 4-CPP; 2,4-D; 3,4-DA; 2,4-DB; 3,4-DB;2,4-DEB; 2,4-DEP; 3,4-DP; 2,3,6-TBA; 2,4,5-T; 2,4,5-TB; acetochlor,acifluorfen, aclonifen, acrolein, alachlor, allidochlor, alloxydim,allyl alcohol, alorac, ametridione, ametryn, amibuzin, amicarbazone,amidosulfuron, aminocyclopyrachlor, aminopyralid, amiprofos-methyl,amitrole, ammonium sulfamate, anilofos, anisuron, asulam, atraton,atrazine, azafenidin, azimsulfuron, aziprotryne, barban, B CPC,beflubutamid, benazolin, bencarbazone, benfluralin, benfuresate,bensulfuron, bensulide, bentazone, benzadox, benzfendizone, benzipram,benzobicyclon, benzofenap, benzofluor, benzoylprop, benzthiazuron,bicyclopyrone, bifenox, bilanafos, bispyribac, borax, bromacil,bromobonil, bromobutide, bromofenoxim, bromoxynil, brompyrazon,butachlor, butafenacil, butamifos, butenachlor, buthidazole, buthiuron,butralin, butroxydim, buturon, butylate, cacodylic acid, cafenstrole,calcium chlorate, calcium cyanamide, cambendichlor, carbasulam,carbetamide, carboxazole chlorprocarb, carfentrazone, CDEA, CEPC,chlomethoxyfen, chloramben, chloranocryl, chlorazifop, chlorazine,chlorbromuron, chlorbufam, chloreturon, chlorfenac, chlorfenprop,chlorflurazole, chlorflurenol, chloridazon, chlorimuron, chlornitrofen,chloropon, chlorotoluron, chloroxuron, chloroxynil, chlorpropham,chlorsulfuron, chlorthal, chlorthiamid, cinidon-ethyl, cinmethylin,cinosulfuron, cisanilide, clethodim, cliodinate, clodinafop, clofop,clomazone, clomeprop, cloprop, cloproxydim, clopyralid, cloransulam,CMA, copper sulfate, CPMF, CPPC, credazine, cresol, cumyluron,cyanatryn, cyanazine, cycloate, cyclosulfamuron, cycloxydim, cycluron,cyhalofop, cyperquat, cyprazine, cyprazole, cypromid, daimuron, dalapon,dazomet, delachlor, desmedipham, desmetryn, di-allate, dicamba,dichlobenil, dichloralurea, dichlormate, dichlorprop, dichlorprop-P,diclofop, diclosulam, diethamquat, diethatyl, difenopenten, difenoxuron,difenzoquat, diflufenican, diflufenzopyr, dimefuron, dimepiperate,dimethachlor, dimethametryn, dimethenamid, dimethenamid-P, dimexano,dimidazon, dinitramine, dinofenate, dinoprop, dinosam, dinoseb,dinoterb, diphenamid, dipropetryn, diquat, disul, dithiopyr, diuron,DMPA, DNOC, DSMA, EBEP, eglinazine, endothal, epronaz, EPTC, erbon,esprocarb, ethalfluralin, ethametsulfuron, ethidimuron, ethiolate,ethofumesate, ethoxyfen, ethoxysulfuron, etinofen, etnipromid,etobenzanid, EXD, fenasulam, fenoprop, fenoxaprop, fenoxaprop-P,fenoxasulfone, fenteracol, fenthiaprop, fentrazamide, fenuron, ferroussulfate, flamprop, flamprop-M, flazasulfuron, florasulam, fluazifop,fluazifop-P, fluazolate, flucarbazone, flucetosulfuron, fluchloralin,flufenacet, flufenican, flufenpyr, flumetsulam, flumezin, flumiclorac,flumioxazin, flumipropyn, fluometuron, fluorodifen, fluoroglycofen,fluoromidine, fluoronitrofen, fluothiuron, flupoxam, flupropacil,flupropanate, flupyrsulfuron, fluridone, fluorochloridone, fluoroxypyr,flurtamone, fluthiacet, fomesafen, foramsulfuron, fosamine, furyloxyfen,glufosinate, glufosinate-P, glyphosate, halosafen, halosulfuron,haloxydine, haloxyfop, haloxyfop-P, hexachloroacetone, hexaflurate,hexazinone, imazamethabenz, imazamox, imazapic, imazapyr, imazaquin,imazethapyr, imazosulfuron, indanofan, indaziflam, iodobonil,iodomethane, iodosulfuron, iofensulfuron, ioxynil, ipazine,ipfencarbazone, iprymidam, isocarbamid, isocil, isomethiozin,isonoruron, isopolinate, isopropalin, isoproturon, isouron, isoxaben,isoxachlortole, isoxaflutole, isoxapyrifop, karbutilate, ketospiradox,lactofen, lenacil, linuron, MAA, MAMA, MCPA, MCPA-thioethyl, MCPB,mecoprop, mecoprop-P, medinoterb, mefenacet, mefluidide, mesoprazine,mesosulfuron, mesotrione, metam, metamifop, metamitron, metazachlor,metazosulfuron, metflurazon, methabenzthiazuron, methalpropalin,methazole, methiobencarb, methiozolin, methiuron, methometon,methoprotryne, methyl bromide, methyl isothiocyanate, methyldymron,metobenzuron, metobromuron, metolachlor, metosulam, metoxuron,metribuzin, metsulfuron, molinate, monalide, monisouron,monochloroacetic acid, monolinuron, monuron, morfamquat, MSMA,naproanilide, napropamide, naptalam, neburon, nicosulfuron,nipyraclofen, nitralin, nitrofen, nitrofluorfen, norflurazon, noruron,OCH, orbencarb, ortho-dichlorobenzene, orthosulfamuron, oryzalin,oxadiargyl, oxadiazon, oxapyrazon, oxasulfuron, oxaziclomefone,oxyfluorfen, parafluoron, paraquat, pebulate, pelargonic acid,pendimethalin, penoxsulam, pentachlorophenol, pentanochlor, pentoxazone,perfluidone, pethoxamid, phenisopham, phenmedipham, phenmedipham-ethyl,phenobenzuron, phenylmercury acetate, picloram, picolinafen, pinoxaden,piperophos, potassium arsenite, potassium azide, potassium cyanate,pretilachlor, primisulfuron, procyazine, prodiamine, profluazol,profluralin, profoxydim, proglinazine, prometon, prometryn, propachlor,propanil, propaquizafop, propazine, propham, propisochlor,propoxycarbazone, propyrisulfuron, propyzamide, prosulfalin,prosulfocarb, prosulfuron, proxan, prynachlor, pydanon, pyraclonil,pyraflufen, pyrasulfotole, pyrazolynate, pyrazosulfuron, pyrazoxyfen,pyribenzoxim, pyributicarb, pyriclor, pyridafol, pyridate, pyriftalid,pyriminobac, pyrimisulfan, pyrithiobac, pyroxasulfone, pyroxsulam,quinclorac, quinmerac, quinoclamine, quinonamid, quizalofop,quizalofop-P, rhodethanil, rimsulfuron, saflufenacil, S-metolachlor,sebuthylazine, secbumeton, sethoxydim, siduron, simazine, simeton,simetryn, SMA, sodium arsenite, sodium azide, sodium chlorate,sulcotrione, sulfallate, sulfentrazone, sulfometuron, sulfosulfuron,sulfuric acid, sulglycapin, swep, TCA, tebutam, tebuthiuron,tefuryltrione, tembotrione, tepraloxydim, terbacil, terbucarb,terbuchlor, terbumeton, terbuthylazine, terbutryn, tetrafluoron,thenylchlor, thiazafluoron, thiazopyr, thidiazimin, thidiazuron,thiencarbazone-methyl, thifensulfuron, thiobencarb, tiocarbazil,tioclorim, topramezone, tralkoxydim, triafamone, tri-allate,triasulfuron, triaziflam, tribenuron, tricamba, triclopyr, tridiphane,trietazine, trifloxysulfuron, trifluralin, triflusulfuron, trifop,trifopsime, trihydroxytriazine, trimeturon, tripropindan, tritactritosulfuron, vemolate, and xylachlor.

Another embodiment of the present disclosure is a method for the controlor prevention of fungal attack. This method comprises applying to thesoil, plant, roots, foliage, seed or locus of the fungus, or to a locusin which the infestation is to be prevented (for example applying tocereal plants), a fungicidally effective amount of one or more of thecompounds of Formula I. The compounds are suitable for treatment ofvarious plants at fungicidal levels, while exhibiting low phytotoxicity.The compounds may be useful both in a protectant and/or an eradicantfashion.

The compounds have been found to have significant fungicidal effectparticularly for agricultural use. Many of the compounds areparticularly effective for use with agricultural crops and horticulturalplants. Additional benefits may include, but are not limited to,improving the health of a plant; improving the yield of a plant (e.g.increased biomass and/or increased content of valuable ingredients);improving the vigor of a plant (e.g. improved plant growth and/orgreener leaves); improving the quality of a plant (e.g. improved contentor composition of certain ingredients); and improving the tolerance toabiotic and/or biotic stress of the plant.

The compositions of Formula I may be effective against pathogen induceddiseases where the plant fungal pathogen belonging to at least onegenera selected from Blumeria, Podosphaera, Sphaerotheca, Uncinula,Erysiphe, Puccinia, Phakopsora, Gymnosporangium, Hemileia, Uromyces,Alternaria, Cercospora, Cladosporium, Cochliobolus, Colletotrichum,Magnaporthe, Mycosphaerella, Phaeosphaeria, Pyrenophora, Ramularia,Rhyncosporium, Septoria, Venturia, Ustilago, Aspergillus, Penicillium,Drechslera, Fusarium, Botrytis, Gibberella, Rhizoctonia,Pseudocercosporella, Sclerotinia, Helminthosporium, Stagonospora,Exserohilum, and Pyricularia. Pathogens such as Venturia inaequalis,Septoria tritici, Cercospora beticola, Cercospora arachidicola,Colletotrichum lagenarium, Puccinia graminis f. sp. tritici, Pucciniarecondita tritici, Uncinula necator, Blumeria graminis, andMycosphaerella fijiensis may be controlled by compositions of Formula I.Additionally, the compositions of Formula I may be effective inpreventing or controlling diseases including apple scab, speckled leafblotch of wheat, leaf spot of sugarbeets, leaf spot of peanut, cucumberanthracnose, wheat leaf rust, grape powdery mildew, wheat powderymildew, and black sigatoka.

The invention provides kits for the treatment or prevention ofagricultural or plant disease or disorders. In one embodiment, the kitincludes a composition containing an effective amount of a compoundherein in a form suitable for delivery to a site plant. In someembodiments, the kit comprises a container which contains a compound ofFormula I; such containers can be boxes, ampules, bottles, vials, tubes,bags, pouches, blister-packs, or other suitable container forms known inthe art. Such containers can be made of plastic, glass, laminated paper,metal foil, or other materials suitable for holding compounds.

If desired the compound(s) of the invention is provided together withinstructions for administering it to a plant, field, or otheragricultural area. The instructions will generally include informationabout the use of the composition for the treatment or prevention of ametalloenzyme-mediated agricultural disease or disorder. In otherembodiments, the instructions include at least one of the following:description of the compound; dosage schedule and administration fortreatment or prevention of a metalloenzyme-mediated agricultural diseaseor disorder; precautions; warnings; description of research studies;and/or references. The instructions may be printed directly on thecontainer (when present), or as a label applied to the container, or asa separate sheet, pamphlet, card, or folder supplied in or with thecontainer.

The compounds of the present disclosure may be effective in use withplants in a disease-inhibiting and phytologically acceptable amount. Theterm “disease-inhibiting and phytologically acceptable amount” refers toan amount of a compound that kills or inhibits the plant disease forwhich control is desired, but is not significantly toxic to the plant.This amount will generally be from about 0.1 to about 1000 ppm (partsper million), with 1 to 500 ppm being preferred. The exact amount of acompound required varies with the fungal disease to be controlled, thetype of formulation employed, the method of application, the particularplant species, climate conditions, and the like. A suitable applicationrate is typically in the range from about 0.10 to about 4 pounds/acre(about 0.01 to 0.45 grams per square meter, g/m²).

Any range or desired value given herein may be extended or alteredwithout losing the effects sought, as is apparent to the skilled personfor an understanding of the teachings herein.

EXAMPLES

The present invention will now be demonstrated using specific examplesthat are not to be construed as limiting.

General Experimental Procedures

Definitions of variables in the structures in schemes herein arecommensurate with those of corresponding positions in the formulaedelineated herein.

Synthesis of Azole Targets

Syntheses of azole targets (Formula I) may be accomplished using theexample synthesis that is shown below (Scheme 1). A broad range ofheterocycles may be prepared starting from functionalized halo-aromaticstarting materials (e.g. A). For the purpose of this example, R₄ is anaryl group further substituted with R₆. R₃ may be part of a fused orunfused bicyclic ring system.

Example 1

1-(5-Chlorothiophen-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(1)

To a stirred solution of 2-iodothiophene (2.5 grams (g), 11.9 millimoles(mmol)) in n-hexane (25 milliliters (mL)) was added N-chlorosuccinimide(NCS; 1.58 g, 11.9 mmol) followed by a catalytic (cat) amount ofperchloric acid (HClO₄) at room temperature (RT), and stirring wascontinued for 24 hours (h) at RT. The reaction mixture was filtered, andthe filtrate was washed with water (H₂O) and brine, dried over anhydroussodium sulfate (Na₂SO₄) and concentrated in vacuo to afford D (1.7 g,6.9 mmol, 58%) as a liquid. ¹H NMR (200 MHz, CDCl₃): δ 7.18 (d, J=4.2Hz, 1 H), 6.69 (d, J=4.2 Hz, 1 H).

To a stirred solution of ethyl 2-bromo-2,2-difluoroacetate (1.6 mL, 13.9mmol) in dimethyl sulfoxide (DMSO; 30 mL) was added copper powder (1.7g, 27.9 mmol) at RT. After stirring for 1 h at RT,2-chloro-5-iodothiophene D (1.7 g, 6.98 mmol) was added, and stirringwas continued for another 12 h at RT. The progress of the reaction wasmonitored by thin layer chromatography (TLC). The reaction was quenchedwith a saturated (satd) ammonium chloride (NH₄Cl) solution and extractedwith dichloromethane (CH₂Cl₂; 3×50 mL). The combined organic layers werewashed with H₂O (2×50 mL) and brine, dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure to afford crude product which uponpurification by column chromatography (eluting with EtOAc/hexane)afforded compound E (0.65 g, 2.7 mmol, 38%) as a liquid. ¹H NMR (200MHz, CDCl₃): δ 7.19-7.17 (m, 1H), 6.89 (d, J=3.8 Hz, 1H), 4.37 (q, J=7.4Hz, 2H), 1.36 (t, J=7.0 Hz, 3H).

To a stirred solution of 1-bromo-2,4-difluorobenzene (0.3 mL, 2.7 mmol)in ether (Et₂O; 20 mL) was added n-butyllithium (n-BuLi, 1.6 M inhexane; 1.77 mL, 2.7 mmol) at −78° C. under an inert atmosphere. Afterbeing stirred for 15 minutes (min) at −78° C., a solution of compound E(0.65 g, 2.7 mmol) in Et₂O (10 mL) was added to the reaction mixture,and stirring was continued for 1 h at −78° C. and for 1 h at RT. Theprogress of the reaction was monitored by TLC. The reaction was quenchedwith a satd NH₄Cl solution and extracted with ethyl acetate (EtOAc; 3×30mL). The combined organic layers were washed with H₂O and brine, driedover anhydrous Na₂SO₄ and concentrated under reduced pressure to affordcrude product which upon purification by column chromatography (elutingwith EtOAc/hexane) afforded compound F (0.5 g, 1.62 mmol, 60%) as asolid. ¹H NMR (200 MHz, CDCl₃): δ 7.90-7.79 (m, 1H), 7.15-7.12 (m, 1H),7.04-6.86 (m, 3H).

To a stirred solution of F (0.5 g, 1.62 mmol) in Et₂O (40 mL) was addedfreshly prepared diazomethane [Nitrosyl methyl urea (NMU; 0.9 g) in 10%potassium hydroxide (KOH; 40 mL)] at 0° C. and the reaction mixture wasthen warmed to RT. After stirring for 2 h at RT, the volatiles wereevaporated under reduced pressure to afford crude product. The crudeproduct was purified by column chromatography (eluting withEtOAc/hexane) to afford epoxide G (0.3 g, 0.93 mmol, 57%) as a solid. ¹HNMR (200 MHz, CDCl₃): δ 7.34-7.27 (m, 1H), 7.09-6.75 (m, 4H), 3.37 (d,J=4.8 Hz, 1H), 2.98 (m, 1H).

To a stirred solution of 1H-tetrazole (0.039 g, 0.55 mmol) inN,N-dimethylformamide (DMF; 5 mL) was added potassium carbonate (K₂CO₃;0.064 g, 0.46 mmol) at RT under a nitrogen (N₂) atmosphere. Afterstirring for 10 min at RT, epoxide G (0.15 g, 0.46 mmol) was added tothe reaction mixture, and the mixture was heated at 65° C. for 8 h. Thereaction mixture was cooled to RT, diluted with H₂O (40 mL) and thenextracted with EtOAc (2×50 mL). The combined organic phases were washedwith H₂O (2×25 mL) and brine (25 mL), dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. The crude material was purified bycolumn chromatography (eluting with EtOAc/hexane) to afford 1 (30 mg,0.13 mmol, 16%) as a solid. ¹H NMR (500 MHz, CDCl₃): δ 8.61 (s, 1H),7.37-7.32 (m, 1H), 6.85 (d, J=3.5 Hz, 1H), 6.80-6.74 (m, 3H), 5.60 (d,J=14.5 Hz, 1H), 5.02 (d, J=14.5 Hz, 1H). HPLC: 94.1%. MS (ESI): m/z 393[M⁺+1].

Compounds 12-16 in Table 1 were prepared using the same conditions ascompound 1 from commercially available starting materials (given inTable 1).

Example 2

1-(4-Bromothiazol-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(2)

Compound 2 was synthesized using the same conditions as compound 1.Yield: 47% (0.022 g). ¹H NMR (200 MHz, CDCl₃): δ 8.73 (s, 1H), 7.35 (m,2H), 6.84-6.74 (m, 2H), 5.66 (d, J=15.0 Hz, 1H), 5.59 (br s, 1H), 5.19(d, J=15.0 Hz, 1H). HPLC: 96.6%. MS (ESI): m/z 438, 440 [M⁺+1)+2].

Example 3

4-(2-(2-(2,4-Difluorophenyl)-1,1-difluoro-2-hydroxy-3-(1H-tetrazol-1-yl)propyl)thiazol-4-yl)benzonitrile(3)

To a suspension of copper powder (1.04 g, 16.46 mmol) in DMSO (20 mL)was added ethyl 2-bromo-2,2-difluoroacetate (1.83 g, 9.0 mmol), and themixture was stirred for 1 h at RT. Then 2,4-dibromothiazole (1.0 g, 4.11mmol) was added, and stirring was continued for 16 h at RT. The progressof the reaction was monitored by TLC. The reaction was quenched withaqueous (aq) NH₄Cl (15 mL) and extracted with CH₂Cl₂ (3×50 mL). Thecombined organic layers were washed with H₂O and brine, dried overanhydrous Na₂SO₄ and concentrated under reduced pressure to afford acrude product. Silica gel column chromatography eluting with 6%EtOAc/hexane afforded the ester (0.35 g, 1.22 mmol, 37%) as a liquid. ¹HNMR (200 MHz, CDCl₃): δ 7.47 (s, 1H), 4.45-4.33 (m, 2H), 1.41-1.33 (m,3H).

To a stirred solution of 1-bromo-2,4-difluorobenzene (0.20 mL, 1.83mmol) in Et₂O (5 mL) was added n-BuLi (2.5 M solution in hexane; 0.7 mL,1.83 mmol) at −78° C., and the mixture was stirred for 30 min. The esterfrom the previous step (0.35 g, 1.22 mmol) in Et₂O (10 mL) was addeddrop-wise and the mixture was stirred for 1 h at −70° C. The temperaturewas raised gradually to ambient temperature, and stirring was continuedfor another 1 h. The reaction mixture was quenched with aq NH₄Cl andextracted with EtOAc (3×20 mL). The combined organic layers were washedwith H₂O and brine, dried over anhydrous Na₂SO₄ and concentrated underreduced pressure. The crude compound was purified by silica gel columnchromatography eluting with 4% EtOAc/Hexane to afford the ketone (0.13g, 0.36 mmol, 30.09%) as a liquid. ¹H NMR (200 MHz, CDCl₃): δ 8.10-8.02(m, 1H), 7.51 (s, 1H), 7.07-6.85 (m, 2H).

To a stirred solution of ketone (0.13 g, 0.36 mmol) in anhydrous Et₂O(30 mL) was added freshly prepared diazomethane [NMU (0.37 g) in 10% KOH(20 mL)] at 0° C., and the mixture was warmed to room temperature. Afterstirring for 1 h at RT, the solvent was evaporated under reducedpressure to afford a crude product. The crude product was purified bysilica gel column chromatography eluting with 4% EtOAc/hexane to affordthe epoxide (0.13 g, 0.36 mmol, 74%) as a liquid. ¹H NMR (200 MHz,CDCl₃): δ 7.51-7.30 (m, 2H), 6.94-6.75 (m, 2H), 3.58 (d, J=5.0 Hz, 1H),3.05-3.03 (m, 1H).

To a stirred solution of epoxide (0.1 g, 0.27 mmol) and4-cyanophenylboronic acid (0.059 g, 0.40 mmol) in tetrahydrofuran(THF)/H₂O (20 mL, 2:1) was added K₂CO₃ (0.112 g, 0.81 mmol) at RT underinert atmosphere. After purging with argon for a period of 10 min,1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II)(Pd(dppf)₂Cl₂; 0.049 g, 0.06 mmol) was added to the reaction mixtureunder argon atmosphere. The resulting mixture was stirred for 16 h at55° C. The reaction mixture was quenched with H₂O and extracted withEtOAc (3×50 mL). The combined organic phases were washed with H₂O andbrine, dried over anhydrous Na₂SO₄ and concentrated. The crude materialwas purified by silica gel column chromatography eluting with 6%EtOAc/hexane to afford the coupled product (0.065 g, 0.16 mmol, 62%) asa solid. ¹H NMR (200 MHz, CDCl₃): δ 7.99 (d, J=6.6 Hz, 2H), 7.81-7.67(m, 3H), 7.67-7.41 (m, 1H), 6.92-6.74 (m, 2H), 3.66 (d, J=5.0 Hz, 1H),3.09-3.07 (m, 1H).

To a stirred solution of the coupled product (0.065 g, 0.16 mmol) in DMF(2 mL) was added 1H-tetrazole (0.013 g, 0.19 mmol) followed by K₂CO₃(0.011 g, 0.08 mmol) at RT under inert atmosphere. The reaction mixturewas stirred for 16 h at 70° C. The reaction mixture was cooled to RT,diluted with H₂O (5 mL) and extracted with EtOAc (2×20 mL). The organiclayer was washed with H₂O and brine and dried over anhydrous Na₂SO₄.After filtering, the solvent was evaporated under reduced pressure togive crude compound. The crude compound was purified by silica gelcolumn chromatography eluting with 30% EtOAc/hexane to afford 3 (15 mg,0.03 mmol, 19%) as a solid. ¹H NMR (500 MHz, CDCl₃): δ 8.71 (s, 1H),7.91 (d, J=8.0 Hz, 2H), 7.78 (s, 1H), 7.77 (d, J=8.5 Hz, 2H), 7.39-7.35(m, 1H), 6.83-6.79 (m, 1H), 6.76-6.73 (m, 1H), 5.87 (s, 1H), 5.67 (d,J=14.5 Hz, 1H), 5.24 (d, J=14.5 Hz, 1H). HPLC: 95.8%. MS (ESI): m/z 461[M⁺+1].

Example 4

1-(6-Chloroquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(4)

To a stirred suspension of copper powder (3.14 g, 47.4 mmol) in DMSO (50mL) was added ethyl 2-bromo-2,2-difluoroacetate (4.99 g, 24.7 mmol) atRT under N₂ atmosphere. After stirring for 1 h at RT,2-bromo-6-chloroquinoline (3.0 g, 12.3 mmol) was then added and stirringwas continued for an additional 16 h at RT. The reaction mixture wasquenched with satd NH₄Cl and extracted with CH₂Cl₂ (3×100 mL). Thecombined organic layers were washed with H₂O and brine, dried overanhydrous Na₂SO₄ and concentrated under reduced pressure. The crudematerial was purified by silica gel column chromatography eluting with3% EtOAc/hexane to afford the 6-chloro-2-quinolinyl ethyl ester (2.6 g,9.12 mmol, 73%) as a solid. ¹H NMR (500 MHz, CDCl₃): δ 8.26 (d, J=8.5Hz, 1H), 8.08 (d, J=9.0 Hz, 1H), 7.87 (s, 1H), 7.82 (d, J=8.5 Hz, 1H),7.71 (dd, J=9.0, 2.0 Hz, 1H), 4.44-4.39 (m, 2H), 1.38-1.34 (m, 3H). MS(ESI): m/z 286 [M⁺+1].

To a stirred solution of 1-bromo-2,4-difluorobenzene (0.15 mL, 1.40mmol) in Et₂O (20 mL) was added n-BuLi (1.6 M in hexane; 0.87 mL, 1.40mmol) at −70° C. under N₂ atmosphere. After being stirred for 15 min at−70° C., the ester (0.4 g, 1.40 mmol) in Et₂O (5 mL) was added toreaction mixture at −70° C. The reaction mixture was stirred for 1 h at0° C., warmed to RT and stirred for another 1 h. The progress of thereaction was monitored by TLC. The reaction was quenched with a satdNH₄Cl solution and extracted with EtOAc (3×10 mL). The combined organiclayers were washed with H₂O and brine, dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. The crude compound was purified bysilica gel column chromatography eluting with 3% EtOAc/hexane to affordthe corresponding ketone (0.35 g, 0.98 mmol, 70%) as a liquid. ¹H NMR(500 MHz, CDCl₃): δ 8.30-8.23 (m, 1H), 8.14-8.07 (m, 1H), 7.96-7.87 (m,2H), 7.76-7.65 (m, 2H), 7.01-6.98 (m, 1H), 6.80-6.76 (m, 1H). MS (ESI):m/z 354, 356 [(M⁺+1)+2].

To a stirred solution of the ketone (0.35 g, 0.98 mmol) in Et₂O (15 mL)was added freshly prepared diazomethane [NMU (0.8 g) in 10% KOH (50 mL)]drop-wise at −5° C. and the mixture was warmed to RT. After stirring for1 h at RT, the volatiles were evaporated under reduced pressure toafford the crude product. The crude product was purified by columnchromatography (1-3% EtOAc/hexane as a gradient) to afford thecorresponding epoxide (0.14 g, 0.68 mmol, 39%) as semi-solid. ¹H NMR(200 MHz, CDCl₃): δ 8.16-8.06 (m, 2H), 7.88-7.85 (m, 1H), 7.74-7.58 (m,2H), 7.43-7.28 (m, 1H), 6.87-6.68 (m, 2H), 3.50 (d, J=5.2 Hz, 1H), 3.01(br s, 1 H). MS (ESI): m/z 368 [M⁺+1].

To a stirred solution of epoxide (0.14 g, 0.38 mmol) in DMF (10 mL) wasadded 1H-tetrazole (0.026 g, 0.38 mmol) followed by K₂CO₃ (0.079 g, 0.57mmol) at RT under inert atmosphere. The reaction mixture was stirred for6 h at 70° C. The reaction mixture was cooled to RT, diluted with H₂O (5mL) and extracted with EtOAc (2×20 mL). The organic layer was washedwith H₂O and brine and dried over anhydrous Na₂SO₄. After filtering, thesolvent was evaporated under reduced pressure to give crude compound.The crude compound was purified by silica gel column chromatographyeluting with 30% EtOAc/hexane to afford 4 (0.085 g, 0.19 mmol, 51%) as awhite solid and1-(6-chloroquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(2H-tetrazol-2-yl)propan-2-ol(16; 0.04 g, 0.09 mmol, 24%) as white solid. ¹H NMR (500 MHz, CDCl₃): δ8.76 (s, 1H), 8.22 (d, J=8.5 Hz, 1H), 8.03 (d, J=9.5 Hz, 1H), 7.87 (s,1H), 7.79 (dd, J=9.0, 2.5 Hz, 1H), 7.67 (d, J=9.0 Hz, 2H), 7.32-7.27 (m,1H), 6.78-6.73 (m, 1H), 6.60-6.57 (m, 1H), 5.64 (d, J=14.5 Hz, 1H), 5.19(d, J=14.5 Hz, 1H). HPLC: 98.6%. MS (ESI): m/z 438 [M⁺+1]. Racemic1-(6-Chloroquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(2H-tetrazol-2-yl)propan-2-ol(minor product, 16): ¹H NMR (500 MHz, CDCl₃): δ 8.26 (s, 1H), 8.20 (d,J=9.0 Hz, 1H), 7.99 (d, J=9.5 Hz, 1H), 7.85 (s, 1H), 7.75 (dd, J=8.5,2.0 Hz, 1H), 7.68 (d, J=8.5 Hz, 1H), 7.41-7.36 (m, 1H), 6.98 (s, 1H),6.81-6.77 (m, 1H), 6.65-6.61 (m, 1H), 5.85 (d, J=14.5 Hz, 1H), 5.49 (d,J=14.5 Hz, 1H). HPLC: 97.8%. MS (ESI): m/z 438 [M⁺+1].

Separation of 4 Enantiomers (+ and −)

The enantiomers of 4 were separated by high performance liquidchromatography (HPLC) using a CHIRALPAK IC® column (250×4.6 mm, 5μ) withmobile phase (A) n-hexane—(B) isopropyl alcohol (IPA) (IsocraticA:B=70:30) and flow rate 1.00 mL/min

Diluent: IPA:Hexane (20:80)

Chiral preparative HPLC separation afforded 4-(−) ([α]_(D)−7.5° (c=0.1%in methyl alcohol (CH₃OH) and 4-(+) ([α]_(D) 3.58° (c=0.1% in CH₃OH).

Compounds 17-33 in Table 1 were prepared using the same conditions ascompound 4 from commercially available starting materials or preparedintermediates (given in Table 1).

Example 5

2-(2,4-Difluorophenyl)-1,1-difluoro-1-(quinolin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(5)

Compound 5 was prepared using the conditions employed for 4 from2-bromoquinoline: 0.020 g isolated as a cream solid. ¹H NMR (500 MHz,CDCl₃): δ 8.78 (s, 1H), 8.31 (d, J=8.5 Hz, 1H), 8.14 (s, 1H), 8.11 (d,J=8.5 Hz, 1H), 7.90-7.85 (m, 2H), 7.71-7.65 (m, 2H), 7.35-7.30 (m, 1H),6.77-6.73 (m, 1H), 6.59-6.55 (m, 1H), 5.68 (d, J=14.0 Hz, 1H), 5.17 (d,J=14.0 Hz, 1H). HPLC: 97.65%. MS (ESI): m/z 404 [M⁺+1].

Example 6

1-(Benzo[d]thiazol-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(6)

Compound 6 was prepared using the conditions employed for 4 from2-bromobenzo[d]thiazole: 0.027 g as a tan solid. ¹H NMR (500 MHz,CDCl₃): δ 8.75 (s, 1H) 8.12 (d, J=8.5 Hz, 1H), 7.95 (d, J=8.0 Hz, 1H),7.63-7.60 (m, 1H), 7.56-7.53 (m, 1H), 6.81-6.76 (m, 1H), 6.70-6.66 (m,1H), 6.42 (s, 1H), 5.73 (d, J=14.5 Hz, 1H), 5.17 (d, J=14.5 Hz, 1H).HPLC: 96.1%. MS (ESI): m/z 410 [M⁺+1].

Example 7

2-(2,4-Difluorophenyl)-1,1-difluoro-1-(pyrimidin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(7)

Compound 7 was prepared using the conditions employed for 1 from2-iodopyrimidine: 0.007 g as a tan solid. ¹H NMR (500 MHz, CDCl₃): δ8.79 (d, J=4.5 Hz, 2H), 8.73 (s, 1H), 7.47-7.45 (m, 1H), 7.36-7.31 (m,1H), 6.79-6.75 (m, 1H), 6.70-6.67 (m, 1H), 6.39 (s, 1H), 5.60 (d, J=14.5Hz, 1H), 5.20 (d, J=14.5 Hz, 1H). HPLC: 98.8%. MS (ESI): m/z 355 [M⁺+1].

Example 8

2-(4-Chloro-2-fluorophenyl)-1-(6-chloroquinolin-2-yl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(8)

Compound 8 was prepared using the conditions employed for 4 from2-bromo-6-chloroquinoline and 1-bromo-2-fluoro-4-chlorobenzene: 0.021 gisolated as a white solid. ¹H NMR (500 MHz, CDCl₃): δ 8.76 (s, 1H), 8.23(d, J=9.0 Hz, 1H), 8.04 (d, J=9.0 Hz, 1H), 7.88 (d, J=2.0 Hz, 1H), 7.79(dd, J=9.0, 2.0 Hz, 1H), 7.72 (s, OH), 7.67 (d, J=9.0 Hz, 1H), 7.28-7.24(m, 1H), 7.04 (dd, J=12.0, 2.0 Hz, 1H), 6.85 (dd, J=8.5, 2.0 Hz, 1H),5.64 (d, J=14.5 Hz, 1H), 5.20 (d, J=14.5 Hz, 1H). HPLC: 99.4%. MS (ESI):m/z 456 [M⁺+1].

Chiral Preparative HPLC Separation of Enantiomers of 8

The enantiomers of 8 (150 mg, 0.33 mmol) were separated by preparativeHPLC using a CHIRALPAK IC® column (250×20 mm, 5μ; mobile phase (A)n-hexane—(B) ethyl alcohol (A:B=90:10) and flow rate 15 mL/min) toobtain 8-(−) (30 mg, 0.066 mmol, 20%) as an off-white solid.

Analytical Data:

Chiral HPLC: 99.88% ee, R_(t)=20.29 min (CHIRALPAK IC® column, 250×4.6mm, 5μ; mobile phase (A) n-hexane—(B) ethyl alcohol (A:B=90:10); flowrate 1.00 mL/min). Optical rotation [α]_(D) ²⁵: −29.44° (c=0.1% inCH₃OH). ¹H NMR (500 MHz, CDCl₃): δ 8.76 (s, 1H), 8.23 (d, J=9.0 Hz, 1H),8.04 (d, J=9.0 Hz, 1H), 7.88 (d, J=2.0 Hz, 1H), 7.79 (dd, J=9.0, 2.0 Hz,1H), 7.72 (s, OH), 7.67 (d, J=9.0 Hz, 1H), 7.28-7.24 (m, 1H), 7.04 (dd,J=12.0, 2.0 Hz, 1H), 6.85 (dd, J=8.5, 2.0 Hz, 1H), 5.64 (d, J=14.5 Hz,1H), 5.20 (d, J=14.5 Hz, 1H). MS (ESI): m/z 454 [M⁺]. HPLC: 99.29%.

Example 9

1-(6-Bromoquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(9)

Compound 9 was prepared using the conditions employed for 4 from2,6-dibromoquinoline: 0.025 g isolated as a yellow solid. ¹H NMR (500MHz, CDCl₃): δ (8.76 (s, 1H), 8.21 (d, J=8.5 Hz, 1H), 8.06 (s, 1H),7.97-7.91 (m, 2H), 7.67-7.65 (m, 2H), 7.32-7.27 (m, 1H), 6.77-6.73 (m,1H), 6.60-6.57 (m, 1H), 5.63 (d, J=14.5 Hz, 1 H), 5.20 (d, J=14.5 Hz,1H). HPLC: 93.3%. MS (ESI): m/z 482, 484 [M⁺, M⁺+2].

Chiral Preparative HPLC Separation of Enantiomers for 9-(+)

The enantiomers of 9 (150 mg, 0.31 mmol) were separated by preparativeHPLC using a CHIRALPAK IC® column (250×20 mm, 5μ) with mobile phase (A)n-hexane—(B) [CH₂Cl₂-ethyl alcohol (80:20)] (A:B=75:25) and flow rate 12mL/min to obtain 9-(+) (30 mg, 0.062 mmol, 20%) as an off-white solid.

Analytical Data:

Chiral HPLC: 99.90% ee, R_(t)=21.25 min (CHIRALPAK IC® column, 250×4.6mm, 5μ; mobile phase (A) n-hexane—(B) ethyl alcohol (A:B=90:10); flowrate 1.00 mL/min). Optical rotation [α]_(D) ²⁵: +5.80° (c=0.1% inCH₃OH). ¹H NMR (500 MHz, CDCl₃): δ 8.76 (s, 1H), 8.21 (d, J=8.5 Hz, 1H),8.06 (d, J=1.5 Hz, 1H), 7.97 (d, J=8.5 Hz, 1H), 7.92 (dd, J=9.0, 1.5 Hz,1H), 7.67 (d, J=9.0 Hz, 1H), 7.66 (br s, OH), 7.32-7.27 (m, 1H),6.77-6.73 (m, 1H), 6.60-6.57 (m, 1H), 5.64 (d, J=14.5 Hz, 1H), 5.20 (d,J=14.5 Hz, 1H). HPLC: 99.55%. MS (ESI): m/z 482 [M⁺].

Example 10

1-(6-Chloroquinoxalin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(10)

Compound 10 was synthesized using conditions employed for 4 from2-bromo-6-chloroquinoxaline. 2-Bromo-6-chloroquinoxaline was prepared inthe following manner. A mixture of 6-chloroquinoxalin-2(1H)-one (1.0 g,5.5 mmol) and phosphorus tribromide (PBr₃; 3.5 mL, 36.1 mmol) was heatedat 120° C. for 4 h. The reaction mass was cooled to RT, diluted withcold H₂O and extracted with CH₂Cl₂. The combined organic extracts weredried over Na₂SO₄, filtered and concentrated under vacuum to get crudeproduct. The crude compound was purified by silica gel columnchromatography (eluting with EtOAc/hexane) to afford2-bromo-6-chloroquinoxaline (550 mg, 2.26 mmol, 42%) as a solid. ¹H NMR(200 MHz, CDCl₃): δ 8.86 (s, 1H), 8.11 (s, 1H), 7.99 (d, J=8.8 Hz, 1H),7.75 (dd, J=9.0, 2.4 Hz, H). MS (ESI): m/z 243 [M⁺].

Compound 10 (25 mg, 0.056 mmol, 26%) was isolated as an off-white solid.¹H NMR (500 MHz, CDCl₃): δ 9.01 (s, 1H), 8.72 (s, 1H), 8.18 (s, 1H),8.02 (d, J=9.0 Hz, 1H), 7.85 (dd, J=9.0, 2.0 Hz, 1H), 7.27 (s, 1H),6.80-6.75 (m, 1H), 6.69-6.64 (m, 1H), 5.78 (s, 1H, OH), 5.70 (d, J=14.5Hz, 1H), 5.20 (d, J=14.5 Hz, 1H). HPLC: 97.9%. MS (ESI): m/z 439 [M⁺+1].

Chiral Preparative HPLC Separation of Enantiomers for 10-(−)

The enantiomers of 10 (70 mg, 0.16 mmol) were separated by preparativeHPLC using a CHIRALPAK IA® column (250×20 mm, 5μ) with mobile phase (A)n-hexane—(B) ethyl alcohol (A:B=70:30) and flow rate 15 mL/min to obtain10-(−) (20 mg, 0.046 mmol, 28%) as an off-white solid.

Analytical Data:

Chiral HPLC: 99.68% ee, R_(t)=10.31 min (CHIRALPAK IA® column, 250×4.6mm, 5μ; mobile phase (A) n-hexane—(B) ethyl alcohol (A:B=85:15); flowrate: 1.00 mL/min). Optical rotation [α]_(D) ²⁵: −18.52° (c=0.1% inCH₃OH). ¹H NMR (500 MHz, CDCl₃): δ 9.01 (s, 1H), 8.72 (s, 1H), 8.18 (d,J=2.0 Hz, 1H), 8.01 (d, J=9.0 Hz, 1H), 7.84 (dd, J=9.0, 2.0 Hz, 1H),7.29-7.26 (m, 1H), 6.81-6.77 (m, 1H), 6.68-6.65 (m, 1H), 5.77 (s, OH),5.71 (d, J=14.5 Hz, 1H), 5.21 (d, J=14.5 Hz, 1H). HPLC: 99.06%. MS(ESI): m/z 439 [M+H]⁺.

Compound 34 in Table 1 was prepared using the same conditions ascompound 10 from commercially available starting materials (given inTable 1).

Example 11

1-(6-Chlorobenzo[d]thiazol-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(11)

Compound 11 was prepared using the conditions employed for 4 from2-bromo-6-chlorobenzo[d]thiazole: 0.017 g isolated as a cream coloredsolid. ¹H NMR (500 MHz, CDCl₃): δ 8.73 (s, 1H), 8.03 (d, J=9.0 Hz, 1H),7.92 (s, 1H), 7.58 (dd, J=9.0, 2.5 Hz, 1H), 7.39-7.34 (m, 1H), 6.81-6.77(m, 1H), 6.71-6.68 (m, 1H), 6.06 (s, 1H), 5.70 (d, J=14.5 Hz, 1H), 5.20(d, J=14.5 Hz, 1H). HPLC: 96.6%. MS (ESI): m/z 444 [M⁺+1].

Example 12 Preparation of Intermediates

2-Bromo-6-(2,2,2-trifluoroethoxy)quinoline (I-A)

To a stirred solution of 2,2,2-trifluoroethanol (10.0 g, 100 mmol) inCH₂Cl₂ (100 mL) were added triethylamine (Et₃N, 27.8 mL, 200 mmol),p-toluenesulfonyl chloride (19.1 g, 100 mmol) and a catalytic amount of4-dimethylaminopyridine (DMAP; 10 mg) at 0° C. under inert atmosphere.The reaction mixture was allowed to warm to RT and stirring wasmaintained for another 5 h. The reaction mixture was diluted with H₂O(100 mL) and extracted with CH₂Cl₂ (3×200 mL). The combined organicextracts were washed with H₂O (50 mL) and brine (50 mL), dried overanhydrous Na₂SO₄ and concentrated under reduced pressure to affordcompound II (25.0 g, 98.42 mmol; crude) as a semi solid. ¹H NMR (200MHz, CDCl₃): δ 7.81 (d, J=8.0 Hz, 2H), 7.38 (d, J=8.0 Hz, 2H), 4.35 (q,J=8.0 Hz, 2H), 2.47 (s, 3H). MS (ESI): m/z 256 [M+2]⁺.

To a stirred solution of 6-hydroxyquinoline (2.0 g, 13.79 mmol) in DMF(15 mL) were added K₂CO₃ (5.71 g, 41.38 mmol) and compound II (7.01 g,27.59 mmol) at RT. The reaction temperature was gradually increased to80° C., at which point the reaction mixture was stirred for another 16h. After completion of reaction (monitored by TLC), the reaction mixturewas cooled to RT, diluted with H₂O (25 mL) and extracted with EtOAc(3×40 mL). The combined organic extracts were washed with brine (30 mL),dried over anhydrous Na₂SO₄ and concentrated under reduced pressure. Thecrude material was purified by silica gel column chromatography (30-35%EtOAc gradient in hexanes) to afford compound I (2.7 g, 11.88 mmol,86%). ¹H NMR (200 MHz, CDCl₃): δ 8.83 (dd, J=4.4, 1.8 Hz, 1H), 8.06-8.03(m, 2H), 7.22-7.18 (m, 2H), 7.05 (d, J=3.0, 1H), 4.50 (q, J=8.0 Hz, 2H).MS (ESI): m/z 228 [M+H]⁺.

To a stirred solution of I (0.6 g, 2.64 mmol) in EtOAc (25 mL) was addedm-chloroperoxybenzoic acid (m-CPBA; 1.14 g, 6.63 mmol) at RT and underinert atmosphere. After completion of reaction (6 h, monitored by TLC),the reaction mixture was quenched with satd sodium bicarbonate (NaHCO₃)solution (30 mL) and extracted with EtOAc (3×40 mL). The combinedorganic extracts were washed with H₂O (30 mL) and brine (30 mL), driedover anhydrous Na₂SO₄ and concentrated under reduced pressure. The crudematerial was purified by silica gel column chromatography (75-85% EtOAcgradient in hexanes) to afford compound J (0.5 g, 2.06 mmol, 77.8%). ¹HNMR (200 MHz, CDCl₃): δ 8.73 (d, J=9.4 Hz, 1H), 8.44 (d, J=6.2 Hz, 1H),7.64 (d, J=8.2 Hz, 1H), 7.46 (dd, J=9.4, 2.8 Hz, 1H), 7.31 (dd, J=8.2,6.2 Hz, 1H), 7.17 (d, J=2.8 Hz, 1H), 4.50 (q, J=8.0 Hz, 2H). MS (ESI):m/z 244 [M+H]⁺.

A stirred solution of quinoline N-oxide J (1.0 g, 4.11 mmol) in aceticanhydride (Ac₂O; 7 mL) was heated at 130-140° C. for 5 h under inertatmosphere. The resultant mixture was cooled to RT, diluted with H₂O (25mL) and extracted with EtOAc (3×30 mL). The combined organic extractswere washed with brine (40 mL), dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. Phosphorus(III) oxybromide (POBr₃;2.95 g, 10.28 mmol) was mixed with the obtained crude material andheated up to 130-140° C., and stirred for 4 h under inert atmosphericconditions. After completion of reaction (monitored by TLC), thereaction mixture was cooled to RT, quenched with ice-cold H₂O (30 mL)and extracted with EtOAc (3×30 mL). The combined organic extracts werewashed with H₂O (30 mL) and brine (30 mL), dried over anhydrous Na₂SO₄and concentrated under reduced pressure. The crude material was purifiedby silica gel column chromatography (10-15% EtOAc gradient in hexanes)to afford 2-bromo-6-(2,2,2-trifluoroethoxy)quinoline (I-A; 0.55 g, 1.79mmol, 43.7%). ¹H NMR (200 MHz, CDCl₃): δ 8.01 (d, J=9.2 Hz, 1H), 7.90(d, J=8.6 Hz, 1H), 7.51 (d, J=8.6 Hz, 1H), 7.42 (dd, J=9.2, 2.8 Hz, 1H),7.10 (d, J=2.8 Hz, 1H), 4.46 (q, J=8.0 Hz, 2H). MS (ESI): m/z 307[M+H]⁺.

2-Bromo-6-(trifluoromethoxy)quinoline (I-B)

To a stirred solution of 4-(trifluoromethoxy)aniline (1.0 g, 5.6 mmol),sodium-3-nitrobenzene sulfonate (1.89 g, 8.4 mmol), boric acid (0.55 g,8.9 mmol) and iron(II) sulfate heptahydrate (FeSO₄.7H₂O; 0.31 g, 1.1mmol) in glycerol (14 mL) was added concentrated sulfuric acid (H₂SO₄;3.4 mL) dropwise at 0° C. The reaction mixture was gradually heated to150° C. and stirred for 5 h. After completion of the reaction (monitoredby TLC), the reaction mixture was poured into ice-cold H₂O (100 mL),made basic with 50% aq sodium hydroxide (NaOH) solution (10 mL) andextracted with Et₂O (4×25 mL). The separated organic layer was washedwith H₂O (50 mL) and brine (50 mL), dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure to obtain the crude material.Purification by silica gel column chromatography (eluting with 12%EtOAc/hexane) afforded the quinoline K (0.95 g, 4.42 mmol, 79%) as acolorless liquid. ¹H NMR (200 MHz, CDCl₃): δ 8.96-8.94 (m, 1H), 8.16 (d,J=8.2 Hz, 2H), 7.65-7.56 (m, 2H), 7.47 (dd, J=8.2, 4.2 Hz, 1H). MS(ESI): m/z 214 [M+H]⁺.

To a stirred solution of K (0.95 g, 4.40 mmol) in EtOAc (10 mL) at 0° C.was added m-CPBA (1.5 g, 8.8 mmol) and the reaction mixture was stirredat RT for 6 h. After completion of the reaction (by TLC), the reactionmixture was quenched with satd NaHCO₃ solution and extracted with EtOAc(2×50 mL). The combined organic extracts were washed with H₂O (20 mL)and brine (20 mL), dried over anhydrous Na₂SO₄ and concentrated underreduced pressure to obtain the crude material. Purification by silicagel column chromatography (eluting with 10% CH₃OH/CH₂Cl₂ afforded L(0.72 g, 3.1 mmol, 70%) as a colorless liquid. ¹H NMR (200 MHz, CDCl₃):δ 8.82 (d, J=9.4 Hz, 1H), 8.55 (d, J=6.2 Hz, 1H), 7.75-7.69 (m, 2H),7.60 (dd, J=9.4, 1.6 Hz, 1H), 7.39 (dd, J=9.4, 6.2 Hz, 1H). MS (ESI):230 [M+H]⁺

A stirred solution of quinoline N-oxide L (1.0 g, 4.3 mmol) in Ac₂O (10mL) was heated at 130-140° C. for 5 h under inert atmosphere. Theresultant mixture was cooled to RT, diluted with H₂O (30 mL) andextracted with EtOAc (3×30 mL). The combined organic extracts werewashed with brine (30 mL), dried over anhydrous Na₂SO₄ and concentratedunder reduced pressure. POBr₃ (2.2 g, 7.7 mmol) was added to theobtained crude material, and the mixture was heated at 130-140° C. for 4h under inert atmospheric conditions. After completion of reaction(monitored by TLC), the reaction mixture was cooled to RT, quenched withice-cold H₂O (50 mL) and extracted with EtOAc (3×50 mL). The combinedorganic extracts were washed with H₂O (50 mL) and brine (50 mL), driedover anhydrous Na₂SO₄ and concentrated under reduced pressure. The crudematerial was purified by silica gel column chromatography (eluting with10-15% EtOAc/hexanes) to afford compound I-B (0.55 g, 1.79 mmol, 43.7%)as a colorless liquid. ¹H NMR (200 MHz, CDCl₃): δ 8.09 (d, J=9.0 Hz,1H), 8.00 (d, J=9.0 Hz, 1H), 7.64-7.59 (m, 3H). MS (ESI): 292 [M+H]⁺.

Quinoline-6-carbonitrile (I-C)

To a stirred solution of 6-bromoquinoline (2.0 g, 9.61 mmol) in pyridine(30 mL) was added CuCN (3.0 g, 33.6 mmol) at RT under inert atmosphere.The reaction mixture was gradually heated to 200° C. and stirred for 8h. After complete consumption of the starting material (by TLC), thereaction mixture was cooled to RT, diluted with ice-cold H₂O (100 mL)and extracted with EtOAc (3×100 mL). The combined organic extracts werewashed with H₂O (50 mL) and brine (50 mL), dried over anhydrous Na₂SO₄and concentrated under reduced pressure to obtain the crude material.Purification by silica gel column chromatography (eluting with 30%EtOAc/hexane) afforded compound I-C (1.25 g, 8.0 mmol, 83%) as whitesolid. ¹H NMR (200 MHz, CDCl₃): δ 9.08-9.05 (m, 1H), 8.25-8.19 (m, 3H),7.86 (dd, J=8.6, 1.8 Hz, 1H), 7.55 (dd, J=8.6, 1.8 Hz, 1H). MS (ESI):155 [M+H]⁺.

6-(Difluoromethyl)quinoline (I-D)

To a stirred solution of quinoline-6-carbaldehyde (200 mg, 1.27 mmol) inCH₂Cl₂ (10 mL) at 0° C. was added (diethylamino)sulfur trifluoride(DAST; 0.2 mL, 1.53 mmol) under inert atmosphere. The reaction mixturewas allowed to warm to RT and stirred for 16 h. After completeconsumption of the starting material (by TLC), the reaction mixture wasdiluted with CH₂Cl₂ (20 mL) and then quenched with satd NaHCO₃ solution(40 mL). The separated organic layer was washed with ice-cold H₂O (20mL) and brine (20 mL), dried over anhydrous Na₂SO₄ and concentratedunder reduced pressure to obtain I-D (100 mg, crude) as a yellow liquid.The product was characterized by ¹H NMR spectroscopy and directly takento the next step without purification. ¹H NMR (200 MHz, CDCl₃): δ 9.00(dd, J=4.2, 1.4 Hz, 1H), 8.25-8.18 (m, 2H), 7.98 (s, 1H), 7.84 (dd,J=8.8, 1.4 Hz, 1H), 7.48 (dd, J=8.8, 4.2 Hz, 1H), 6.84 (t, J_(F,H)=74.0Hz, 1H). MS (ESI): 180 [M+H]⁺.

5-(2,2,2-Trifluoroethoxy)quinoline (I-E)

To a stirred solution of amine 5-aminoquinoline (5.0 g, 34.67 mmol) inH₂O (100 mL) was added sodium bisulfate (NaHSO₃; 25.2 g, 242.1 mmol) atRT, and the mixture was stirred at reflux temperature for 36 h. Theresulting solution was cooled to RT, NaOH (9.7 g, 242.5 mmol) was added,and the mixture was stirred at reflux temperature for 8 h. Aftercompletion of the reaction (monitored by TLC), the reaction mixture wascooled to RT and the pH was adjusted to 7.0 with 6 Normal (N)hydrochloride acid (HCl). The precipitate was filtered, washed with H₂O,and dried under high vacuum to obtain the desired alcohol M (3.2 g,22.04 mmol, 64%) as pale-yellow solid. ¹H NMR (500 MHz, CDCl₃): δ 8.92(s, 1H), 8.58 (d, J=8.5 Hz, 1H), 7.71 (d, J=8.5 Hz, 1H), 7.53 (t, J=8.0Hz, 1H), 7.41 (dd, J=8.5, 4.5 Hz, 1H), 6.88 (d, J=7.5 Hz, 1H), 6.10 (brs, 1H). MS (ESI): m/z 146 [M+H]⁺.

A stirred solution of alcohol M (3.2 g, 22.04 mmol),2,2,2-trifluoroethyl-4-methylbenzenesulfonate (5.6 g, 22.04 mmol) andK₂CO₃ (9.12 g, 66.08 mmol) in DMF (40 mL) was heated at 120° C. under aninert atmosphere. After completion of reaction (16 h, monitored by TLC),the reaction mixture was cooled to RT, diluted with H₂O (30 mL) and thenextracted with EtOAc (3×40 mL). The combined organic extracts werewashed with brine (40 mL), dried over anhydrous Na₂SO₄ and concentratedunder reduced pressure to obtain the crude material. Purification bysilica gel column chromatography (eluting with a 3-8% EtOAc gradient inhexanes) provided compound I-E (3.2 g, 14.08 mmol, 63.9%) as a whitesolid. ¹H NMR (500 MHz, CDCl₃): δ 8.94 (d, J=6.0 Hz, 1H), 8.59 (d, J=8.5Hz, 1H), 7.80 (d, J=9.0 Hz, 1H), 7.63-7.60 (m, 1H), 7.44 (dd, J=8.5, 4.5Hz, 1H), 6.86 (d, J=8.0 Hz, 1H), 4.57-4.52 (m, 2H). MS (ESI): m/z 228.0[M+H]⁺.

4,4,5,5-Tetramethyl-2-(4-(2,2,2-trifluoroethoxy)phenyl)-1,3,2-dioxaborolane(I-F)

To a stirred solution of boronate ester N (300 mg, 1.36 mmol) in DMF (10mL) was added K₂CO₃ (940 mg, 6.81 mmol) followed by compound II (342 mg,1.36 mmol) at RT under inert atmosphere. The reaction mixture wasgradually heated to 120° C. and stirred for 24 h. Progress of thereaction was monitored by TLC. The reaction mixture was allowed to coolto RT and diluted with H₂O (50 mL). The aqueous layer was extracted withEtOAc (2×50 mL). The combined organic layer was washed with H₂O (50 mL)and brine (50 mL), dried over anhydrous Na₂SO₄ and concentrated invacuo. The crude material was purified by silica gel columnchromatography (eluting with 4% EtOAc/Hexane) to afford I-F (40 mg, 13.2mmol, 9.7%) as a semi-solid. ¹H NMR (200 MHz, CDCl₃): δ 7.78 (d, J=8.5Hz, 2H), 6.93 (d, J=8.5 Hz, 2H), 4.37 (q, J=8.2 Hz, 2H), 1.36 (s, 12H).

I-F may also be synthesized in a two-step procedure. To a stirredsolution of p-bromo phenol (1.5 g, 8.67 mmol) in DMF (15 mL) was addedK₂CO₃ (6.0 g, 43.3 mmol) followed by tosyl compound II (2.2 g, 8.67mmol) at RT under inert atmosphere. The reaction mixture was graduallyheated to 110° C. for 4 h. After complete consumption of the startingmaterial (by TLC), the reaction mixture was allowed to cool to RT,diluted with H₂O (100 mL) and the aqueous layer was extracted with EtOAc(2×100 mL). The combined organic layer was washed with H₂O (50 mL) andbrine (50 mL), dried over anhydrous Na₂SO₄ and concentrated in vacuo.The crude material was purified by silica gel column chromatography(eluting with 3% EtOAc/hexane) to afford O (1.7 g, 6.66 mmol, 76%) assemi-solid. ¹H NMR (200 MHz, CDCl₃): δ 7.45-7.38 (m, 2H), 6.87-6.79 (m,2H), 4.32 (q, J=8.2 Hz, 2H).

To a stirred solution of O (0.5 g, 1.96 mmol) in 1,4-dioxane (50 mL) wasadded bis(pinacolato) diboron (0.54 g, 2.15 mmol) followed by potassiumacetate (KOAc; 0.576 g, 5.88 mmol) at RT under inert atmosphere. Afterpurging with N₂ for 10 min, Pd(dppf)₂Cl₂ (72 mg, 0.09 mmol) was added tothe reaction mixture under N₂ atmosphere. The mixture was graduallyheated to 110° C. and stirred for 2 h. After complete consumption of thestarting material (by TLC), the volatiles were evaporated under reducedpressure; the obtained residue was dissolved in H₂O (100 mL) andextracted with EtOAc (3×50 mL). The combined organic extracts werewashed with H₂O (50 mL) and brine (50 mL), dried over anhydrous Na₂SO₄and concentrated in vacuo. The crude material was purified by silica gelcolumn chromatography (eluting with 3-4% EtOAc/hexane) to afford I-F(0.28 g, 0.92 mmol, 47%) as a yellow syrup. ¹H NMR (200 MHz, CDCl₃): δ7.78 (d, J=8.5 Hz, 2H), 6.93 (d, J=8.5 Hz, 2H), 4.37 (q, J=8.2 Hz, 2H),1.36 (s, 12H).

Example 13

1-(6-Chloroquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(4H-1,2,4-triazol-4-yl)propan-2-ol(35)

To a stirred suspension of copper powder (2.6 g, 41.23 mmol) in DMSO (25mL) was added ethyl 2-bromo-2,2-difluoroacetate (2.6 mL, 20.62 mmol) atRT under N₂ atmosphere. After being stirred for 1 h at RT,2-bromo-6-chloroquinoline (2.5 g, 10.31 mmol) was added to the reactionmixture, and stirring was continued for another 16 h at RT. Aftercompletion of reaction (by TLC), the reaction mixture was quenched withsatd NH₄Cl solution and extracted with EtOAc (2×200 mL). The combinedorganic extracts were washed with H₂O (50 mL) and brine (50 mL), driedover anhydrous Na₂SO₄ and concentrated under reduced pressure to obtainthe crude material. Purification by silica gel column chromatography(eluting with 3% EtOAc/Hexane) afforded P (2.7 g, 9.47 mmol, 91%) as awhite solid. ¹H NMR (500 MHz, CDCl₃): δ 8.26 (d, J=8.5 Hz, 1H), 8.08 (d,J=9.0 Hz, 1H), 7.87 (s, 1H), 7.82 (d, J=8.5 Hz, 1H), 7.71 (dd, J=9.0,2.0 Hz, 1H), 4.44-4.39 (m, 2H), 1.38-1.34 (m, 3H). MS (ESI): m/z 286[M+1]⁺.

To a stirred solution of 1-bromo-2,4-difluorobenzene (0.6 mL, 4.89 mmol)in Et₂O (20 mL) was added n-BuLi (2.5 M in hexane; 2 mL, 4.89 mmol) at−78° C. under N₂ atmosphere. After being stirred for 15 min, ester P(0.7 g, 2.44 mmol) in Et₂O (10 mL) was added at −78° C., and stirringwas continued for another 2 h. The progress of the reaction wasmonitored by TLC. The reaction was quenched with satd NH₄Cl solution andextracted with CH₂Cl₂ (2×50 mL). The combined organic extracts werewashed with H₂O (50 mL) and brine (50 mL), dried over anhydrous Na₂SO₄and concentrated under reduced pressure to afford ketone Q (0.5 g,crude). The crude mixture was taken to the next step withoutpurification. ¹H NMR (500 MHz, CDCl₃): δ 8.30-8.23 (m, 1H), 8.14-8.07(m, 1H), 7.96-7.87 (m, 2H), 7.76-7.65 (m, 2H), 7.01-6.98 (m, 1H),6.80-6.76 (m, 1H). MS (ESI): m/z 354 [M+1]⁺.

To a stirred solution of ketone Q (0.5 g, crude) in Et₂O (50 mL) wasadded freshly prepared diazomethane [prepared by dissolving NMU (2 g) in1:1 mixture of 10% KOH solution (50 mL) and Et₂O (50 mL) at −5° C.followed by separation and drying of the organic layer using KOHpellets]. After being stirred for 1 h at RT, the volatiles wereevaporated under reduced pressure to obtain the crude material. Thecrude product was purified by silica gel column chromatography (elutingwith 1-3% EtOAc/Hexane) to afford compound R (0.35 g, 0.95 mmol) aswhite solid. ¹H NMR (200 MHz, CDCl₃): δ 8.16-8.06 (m, 2H), 7.88-7.85 (m,1H), 7.74-7.58 (m, 2H), 7.43-7.28 (m, 1H), 6.87-6.68 (m, 2H), 3.50 (d,J=5.2 Hz, 1H), 3.03-3.01 (m, 1H). MS (ESI): m/z 368 [M+1]⁺.

To a stirred solution of epoxide R (300 mg, 0.817 mmol) in DMF (5 mL)was added aq ammonia (NH₃; 5 mL) at RT. The reaction mixture wasgradually heated up to 80° C. and stirred for 3 h. The progress of thereaction was monitored by TLC. The volatiles were evaporated underreduced pressure, and the residue was diluted with EtOAc. The organiclayer was then washed with H₂O (25 mL) and brine (25 mL), dried overanhydrous Na₂SO₄ and concentrated under reduced pressure to obtain thecrude material. Purification by silica gel column chromatography(eluting with 3% CH₃OH/CH₂Cl₂) afforded amine S (100 mg, 0.26 mmol, 31%)as a syrup. ¹H NMR (200 MHz, CDCl₃): δ 8.14-8.01 (m, 2H), 7.85 (s, 1H),7.70 (dd, J=9.0, 2.2 Hz, 1H), 7.58-7.45 (m, 2H), 6.82-6.70 (m, 2H), 3.85(dd, J=14.0, 4.8 Hz, 1H), 3.30 (d, J=14.0 Hz, 1H). MS (ESI): m/z 385[M+1]⁺.

To a stirred solution of N-formyl hydrazine (16 mg, 0.26 mmol) in CH₃OH(5 mL) was added triethyl orthoformate (0.1 mL, 0.26 mmol) at RT underinert atmosphere. The reaction mixture was then heated at 80° C. for 3h; progress of the reaction was monitored by TLC. The reaction mixturewas cooled to 40° C., S (50 mg, 0.13 mmol) was added and stirring wascontinued for another 3 h at 80° C. The volatiles were evaporated underreduced pressure, and the residue was diluted with EtOAc (25 mL). Theorganic layer was washed with H₂O (25 mL) and brine (25 mL), dried overNa₂SO₄ and concentrated under reduced pressure to obtain the crudeproduct. The crude material was purified by silica gel columnchromatography (eluting with 7% CH₃OH/CH₂Cl₂) to afford 35 (25 mg, 0.05mmol, 44%) as colorless semi-solid. ¹H NMR (500 MHz, CDCl₃): δ 8.21 (d,J=9.0 Hz, 1H), 8.16 (s, 2H), 8.08 (d, J=9.0 Hz, 1H), 7.87 (s, 1H),7.82-7.78 (m, 2H), 7.64 (d, J=8.5 Hz, 1H), 7.32-7.29 (m, 1H), 6.77-6.72(m, 1H), 6.58-6.55 (m, 1H), 5.14 (d, J=14.0 Hz, 1H), 4.75 (d, J=14.0 Hz,1H). HPLC: 85.3%. MS (ESI): m/z 437 [M+1]⁺.

Example 14

2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-3-yl)propan-2-ol(36)

To a stirred solution of 2,5-dibromopyridine (5.0 g, 21.09 mmol) in Et₂O(200 mL) was added n-BuLi (2.4 M in hexane; 10.5 mL, 25.3 mmol) at −78°C., and the reaction mixture was stirred for 1 h under inert atmosphere.Diethyl oxalate (4.0 mL, 25.3 mmol) was added to the reaction mixture at−78° C., and stirring was continued for another 10 min. The reactionmixture was allowed to warm to 0° C. and stirred for 2 h. After completeconsumption of the starting material (by TLC), the reaction mixture wasquenched with satd NH₄Cl solution and extracted with CH₂Cl₂ (2×100 mL).The combined organic extracts were washed with H₂O (40 mL) and brine (40mL), dried over anhydrous Na₂SO₄ and concentrated under reduced pressureto obtain the crude. The crude material was purified by silica gelcolumn chromatography (eluting with 8% EtOAc/hexane) to afford compoundT (0.87 g, 3.37 mmol 16%) as a pale-yellow liquid. ¹H NMR (500 MHz,CDCl₃): δ 9.09 (s, 1H), 8.34 (d, J=7.5 Hz, 1H), 7.49 (d, J=8.5 Hz, 1H),4.46 (q, J=7.0 Hz, 2H), 1.44 (t, J=7.0 Hz, 3H). MS (ESI): m/z 259.2[M+H]⁺.

To a stirred solution of compound T (180 mg, 0.69 mmol) in CH₂Cl₂ (10mL) was added DAST (140 mg, 0.87 mmol) at 0° C. under inert atmosphere.The resultant reaction mixture was allowed to warm to RT and stirred for16 h. After complete consumption of the starting material (by TLC), thereaction mixture was diluted with CH₂Cl₂ (50 mL) and washed withice-cold H₂O (50 mL) and brine (50 mL), dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure to obtain the crude material.Purification by silica gel column chromatography (eluting with 6%EtOAc/hexane) afforded ester U (105 mg, 0.37 mmol, 54%) as a yellowliquid. ¹H NMR (500 MHz, CDCl₃): δ 8.62 (d, J=2.0 Hz, 1H), 7.77 (dd,J=8.0 Hz, 2.0 Hz, 1H), 7.60 (d, J=8.0 Hz, 1H), 4.33 (q, J=7.0 Hz, 2H),1.29 (t, J=7.0 Hz, 3H). MS (ESI): m/z 282 [M+2]⁺.

To a stirred solution of 1-bromo-2,4-difluorobenzene (0.1 mL, 0.38 mmol)in Et₂O (5 mL) was added n-BuLi (2.3 M in hexane; 0.16 mL, 0.38 mmol) at−78° C., and the reaction mixture was stirred for 30 min under inertatmosphere. A solution of ester U (90 mg, 0.32 mmol) in Et₂O (5 mL) wasadded to the reaction mixture at −78° C., and stirring was continued foranother 2 h. After complete consumption of the starting material (byTLC), the reaction was quenched with satd NH₄Cl solution, and thereaction mixture was extracted with EtOAc (2×30 mL). The combinedorganic extracts were washed with H₂O (30 mL) and brine (30 mL), driedover anhydrous Na₂SO₄ and concentrated under reduced pressure to affordketone V (0.37 g, crude). This was used in the next step without anyfurther purification. ¹H NMR (500 MHz, CDCl₃): δ 8.60 (s, 1H), 7.91-7.87(m, 1H), 7.77-7.75 (m, 1H), 7.62 (d, J=7.0 Hz, 1H), 7.05-7.01 (m, 1H),6.93-6.88 (m, 1H). MS (ESI): m/z 348 [M+H]⁺.

To a stirred solution of ketone V (80 mg, crude) in Et₂O (10 mL) wasadded freshly prepared diazomethane [prepared by dissolving NMU (200 mg,2.06 mmol) in a 1:1 mixture of 10% KOH solution (20 mL) and Et₂O (20 mL)at 0° C. followed by separation and drying of the organic layer usingKOH pellets] at −5° C., and the reaction mixture was stirred for 2 h.The resulting reaction mixture was allowed to warm to RT, and stirringwas continued for another 16 h. After complete consumption of thestarting material (by TLC), the reaction mixture was concentrated underreduced pressure to obtain crude epoxide W (54 mg). The crude productwas confirmed by ¹H-NMR analysis and was used in the next reactionwithout further purification. ¹H NMR (500 MHz, CDCl₃): δ 8.37 (s, 1H),7.57-7.52 (m, 2H), 7.28-7.2 (m, 1H), 6.87-6.78 (m, 2H), 3.28 (d, J=5.0Hz, 1H), 2.97-2.91 (m, 1H). MS (ESI): m/z 364 [M+2]⁺.

To a stirred solution of epoxide W (120 mg, 0.33 mmol) in THF/H₂O (20mL, 8:2) was added K₂CO₃ (137 mg, 0.99 mmol) followed by boronate I-F(110 mg, 0.363 mmol) at RT under inert atmosphere. After purging with N₂for 45 min, Pd(dppf)₂Cl₂ (12 mg, 0.016 mmol) was added to the reactionmixture under an inert atmosphere, and the resulting mixture was stirredat 70° C. for 2 h. After complete consumption of the starting material(by TLC), the reaction mixture was allowed to cool to RT, diluted withH₂O (100 mL) and the aqueous layer was extracted with EtOAc (2×200 mL).The combined organic extracts were washed with H₂O (100 mL) and brine(100 mL), dried over anhydrous Na₂SO₄ and concentrated in vacuo. Thecrude material was purified by silica gel column chromatography (elutingwith 5-6% EtOAc/hexane) to afford X (115 mg, 0.25 mmol, 75%) as a whitesolid. ¹H NMR (200 MHz, CDCl₃): δ 8.63 (s, 1H), 8.03 (d, J=8.5 Hz, 2H),7.75-7.69 (m, 2H), 7.31-7.28 (m, 1H), 7.06 (d, J=9.0 Hz, 2H), 6.86-6.83(m, 1H), 6.81-6.77 (m, 1H), 4.42 (q, J=8.2 Hz, 2H), 3.32 (d, J=5.0 Hz,1H), 2.98-2.97 (m, 1H). MS (ESI): m/z 456 [M−H]⁻.

To a stirred solution of epoxide X (115 mg, 0.25 mmol) in dry DMF (10mL) was added 1H-tetrazole (28 mg, 0.37 mmol) followed by K₂CO₃ (52 mg,0.25 mmol) at RT under inert atmosphere. The reaction mixture wasgradually heated up to 65° C. and stirred for 20 h. After completeconsumption of the starting material (by TLC), the reaction mixture wasdiluted with ice-cold H₂O (30 mL) and extracted with EtOAc (2×50 mL).The separated organic layer was washed with H₂O (30 mL) and brine (30mL), dried over anhydrous Na₂SO₄ and concentrated under reduced pressureto obtain crude product. The crude material was purified by silica gelcolumn chromatography (eluting with 45% EtOAc/Hexane) to afford 36 (48mg, 0.09 mmol, 36%) as an off-white solid. ¹H NMR (500 MHz, CD₃OD): δ8.66 (s, 1H), 8.42 (s, 1H), 7.99 (d, J=7.5 Hz, 2H), 7.63 (d, J=7.5 Hz,1H), 7.55 (d, J=7.0 Hz, 1H), 7.10-7.03 (m, 3H), 6.81-6.79 (m, 1H),6.68-6.64 (m, 1H), 5.73 (d, J=14.5 Hz, 1H), 5.14 (d, J=14.5 Hz, 1H),4.44 (q, J=8.2 Hz, 2H), 4.35 (s, OH). HPLC: 96.1%. MS (ESI): m/z 528[M+H]⁺.

Example 15

1-(7-Chloroisoquinolin-3-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(37)

To a stirred solution of 2-(methylamino)acetonitrile hydrochloride (4.17g, 39.13 mmol) in CH₃OH/H₂O (45 mL, 2:1 v/v) was added NaCN (2.1 g,42.68 mmol), and the reaction mixture was maintained at RT for 5 min. Tothe above solution, 4-chlorobenzaldehyde (5.0 g, 35.56 mmol) in CH₃OH(30 mL) was added slowly dropwise at the same temperature for 20 min,then the temperature was gradually increased to 70° C. and maintainedfor 8 h. After completion of reaction (monitored by TLC, 20%EtOAc/hexanes), the reaction was quenched with H₂O (20 mL) and extractedwith EtOAc (3×100 mL). The combined extracts were washed with H₂O (25mL), brine (25 mL), dried over anhydrous Na₂SO₄ and concentrated underreduced pressure. The crude product was purified by silica gel columnchromatography (eluting with 15-20% EtOAc gradient in hexanes) to yieldY (4.0 g, 18.21 mmol, 51%) as a thick syrup. ¹H NMR (500 MHz, CDCl₃): δ7.48 (d, J=8.7 Hz, 2H), 7.43 (d, J=8.7 Hz, 2H), 4.86 (s, 1H), 3.47 (d,J=14.0 Hz, 1H), 3.45 (d, J=14.0 Hz, 1H), 2.51 (s, 3H). MS (ESI): m/z 218[M−H]⁻.

A neat mixture of cyano compound Y (5.0 g, 22.76 mmol) indimethylsulfate (8.6 mL, 91.04 mmol) was heated at 120° C. under inertconditions for 6 h. After completion of reaction (monitored by TLC, 30%EtOAc/hexanes), the reaction mixture was cooled to RT and directly takenon to the next step without any purification.

To a stirred solution of methyl sulfate salt Z (5.0 g, crude) in CH₂Cl₂(50 mL) was added aq NH₃ solution (40 mL) under inert atmosphere at −25°C., and the reaction mixture was maintained for 30 min under the sameconditions. After completion of reaction (monitored by TLC, 30%EtOAc/hexanes), the reaction mixture was diluted with H₂O (40 mL) andextracted with CH₂Cl₂ (3×50 mL). The combined organic extracts weredried over anhydrous Na₂SO₄ and concentrated under reduced pressure toobtain the crude AA. ¹H NMR (500 MHz, CDCl₃): δ 7.67 (s, 1H), 7.43-7.42(m, 2H), 4.92 (s, 1H), 3.94 (s, 2H), 2.30 (s, 6H). MS (ESI): m/z 234[M+H]⁺.

The obtained crude product AA (˜5.0 g) was dissolved in ethyl alcohol(EtOH; 40 mL), and stirring was maintained under inert atmosphere. Tothis stirred solution was added copper(II) sulfate pentahydrate(CuSO₄.5H₂O; 7.5 g, 30.03 mmol) in H₂O (40 mL) at RT, and the mixturewas gently heated up to reflux for 30 min. After completion of reaction(monitored by TLC, 30% EtOAc/hexanes), the reaction mixture was cooledto RT and then filtered. The filtrate was extracted with CH₂Cl₂ (2×70mL); the combined extracts were washed with brine (30 mL), dried overanhydrous Na₂SO₄ and concentrated under reduced pressure to obtain thecrude material. Purification by silica gel column chromatography(eluting with 15-20% EtOAc gradient in hexanes) afforded AB (1.2 g, 6.68mmol, 29% overall yield from Y in three sequential steps). ¹H NMR (500MHz, CDCl₃): δ 10.03 (s, 1H), 7.82 (d, J=2.0 Hz, 1H), 7.65-7.62 (m, 2H),4.23 (s, 2H). MS (ESI): m/z 178 [M−H]⁻.

To a stirred solution of benzaldehyde AB (2.0 g, 11.13 mmol) in EtOH (25mL) were added 4-methoxybenzylamine (PMBNH₂; 1.91 g, 13.92 mmol) and acatalytic amount of trifluoroacetic acid (TFA; 5 mol %) at RT, and themixture was gradually heated to reflux temperature under inertatmosphere. After completion of reaction (8 h, monitored by TLC), thereaction mixture was cooled to RT and concentrated under reducedpressure. The obtained solid residue was subjected to crystallization(50% CH₂Cl₂/pentane) to afford colorless crystalline isoquinolinederivative AC (2.0 g, 6.69 mmol, 60%). ¹H NMR (500 MHz, CDCl₃): δ 8.75(s, 1H), 7.72 (s, 1H), 7.45 (d, J=8.5 Hz, 1H), 7.38 (d, J=8.5 Hz, 1H),7.32 (d, J=9.0 Hz, 2H), 6.89 (d, J=9.0 Hz, 2H), 6.46 (s, 1H), 5.02 (brs, 1H), 4.43 (s, 2H), 3.80 (s, 3H). MS (ESI): m/z 299 [M+H]⁺.

To a stirred solution of compound AC (2.0 g, 6.69 mmol) in CH₂Cl₂ (40mL) was added TFA (20 mL) at RT and refluxed for 3 h under inertcondition. After completion of reaction (monitored by TLC), the reactionmixture was cooled to RT, quenched with satd NaHCO₃ solution (40 mL) andextracted with CH₂Cl₂ (3×40 mL). The combined extracts were washed withbrine (25 mL), dried over anhydrous Na₂SO₄ and evaporated in vacuo. Theobtained crude material was purified by silica gel column chromatography(eluting with EtOAc/hexane) to afford amine AD (1.11 g, 6.21 mmol, 93%)as a yellowish solid. ¹H NMR (500 MHz, CDCl₃): δ 8.78 (s, 1H), 7.75 (s,1H), 7.48 (d, J=9.0 Hz, 1H), 7.42 (d, J=9.0 Hz, 1H), 6.70 (s, 1H), 4.48(br s, 2H). MS (ESI): m/z 179 [M+H]⁺.

To a stirred solution of amine AD (1.0 g, 5.60 mmol) in 48% aqhydrobromic acid (HBr) solution (4.8 mL) was added sodium nitrite(NaNO₂; 0.58 g, 8.40 mmol) in H₂O (20 mL) dropwise at 0° C. for 15 min,and the reaction mixture was maintained at the same temperature for 1 h.After completion of reaction (monitored by TLC, 40% EtOAc/hexanes), thereaction mixture was diluted with H₂O (20 mL), made basic (pH 8-9) with2 N aq NaOH solution and then extracted with Et₂O (3×30 mL). Thecombined organic extracts were washed with brine (30 mL), dried overanhydrous Na₂SO₄ and evaporated in vacuo. The crude material waspurified by silica gel column chromatography (eluting with 5-10% EtOAcgradient in hexanes) to afford compound AE (0.3 g, 1.23 mmol, 22%) as anoff-white solid. ¹H NMR (500 MHz, CDCl₃): δ 8.97 (s, 1H), 7.95 (s, 1H),7.89 (s, 1H), 7.72 (d, J=8.5 Hz, 1H), 7.66 (d, J=8.5 Hz, 1H). MS (ESI):m/z 242 [M⁺].

To a stirred suspension of Copper-Bronze (1.52 g, 8.25 mmol) in DMSO (10mL) was added ethyl 2-bromo-2,2-difluoroacetate (0.55 mL, 4.13 mmol) atRT. After being stirred at RT for 1 h, compound AE (0.5 g, 2.06 mmol)was added portionwise, and the mixture was stirred for 16 h under inertatmosphere. After completion of reaction (by TLC), the reaction mixturewas quenched with satd NH₄Cl solution (50 mL), filtered through aCelite® pad and washed with CH₂Cl₂ (3×50 mL). The separated organiclayer was washed with brine (50 mL), dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. The crude material was purified bysilica gel column chromatography (eluting with 5-10% EtOAc in hexanes)to afford ester AF (0.25 g, 0.87 mmol, 42%) as a thick syrup. ¹H NMR(500 MHz, CDCl₃): δ 9.20 (s, 1H), 8.12 (s, 1H), 8.03 (s, 1H), 7.91 (d,J=8.5 Hz, 1H), 7.75 (d, J=8.5 Hz, 1H), 4.38 (q, J=7.0 Hz, 2H), 1.33 (t,J=7.0 Hz, 3H). MS (ESI): m/z 286 [M+H]⁺.

To a stirred solution of 1-bromo-2,4-difluorobenzene (68 mg, 0.35 mmol)in Et₂O (6 mL) was added n-BuLi (1.6 M in hexanes; 0.22 mL, 0.35 mmol)dropwise at −78° C. under inert atmosphere, and the mixture was stirredfor 20 min. A solution of ester AF (100 mg, 0.35 mmol) in Et₂O (5 mL)was added to the reaction mixture at −78° C., and stirring was continuedat the same temperature for 1 h and then at RT for 15 min. Progress ofthe reaction was monitored by TLC. The reaction was quenched with satdNH₄Cl solution, and the mixture was extracted with EtOAc (2×25 mL). Thecombined organic extracts were washed with H₂O (20 mL) and brine (20mL), dried over anhydrous Na₂SO₄ and concentrated under reduced pressureto afford ketone AG (100 mg, crude). The crude material was taken on tothe next step without further purification. MS (ESI): m/z 354 [M+H]⁺.

To a stirred solution of ketone AG (500 mg, crude) in Et₂O (25 mL) wasadded freshly prepared diazomethane [prepared by dissolving NMU (1.0 g,9.71 mmol) in a mixture of 10% aq KOH solution (50 mL) and Et₂O (25 mL)at 0° C. followed by separation and drying of the organic layer usingKOH pellets] at 0° C., and the reaction mixture was maintained at 0°C.—RT for 4 h. The progress of the reaction was monitored by TLC (20%EtOAc/hexanes). The reaction mixture was concentrated under reducedpressure to afford crude product. The crude material was purified bysilica gel column chromatography (eluting with 10-15% EtOAc gradient inhexanes) followed by preparative HPLC to afford the epoxide AH (70 mg,0.19 mmol, 13% from ester AF in two steps). ¹H NMR (500 MHz, CDCl₃): δ9.21 (s, 1H), 8.02 (s, 1H), 7.85 (s, 1H), 7.82 (d, J=9.0 Hz, 1H), 7.71(d, J=9.0 Hz, 1H), 7.43-7.26 (m, 1H), 6.79-6.62 (m, 1H), 6.72-6.68 (m,1H), 3.53 (d, J=5.0 Hz, 1H), 3.01 (d, J=5.0 Hz, 1H). HPLC: 99.9%. MS(ESI): m/z 368 [M+H]⁺.

To a stirred solution of epoxide AH (70 mg, 0.19 mmol) in dry DMF (5 mL)was added 1H-tetrazole (20 mg, 0.28 mmol) followed by K₂CO₃ (26.3 mg,0.19 mmol) at RT under inert atmosphere. The resulting reaction mixturewas gradually heated up to 65° C. and stirred for 16 h; progress of thereaction was monitored by TLC. The reaction mixture was then dilutedwith ice-cold H₂O and extracted with EtOAc (2×30 mL). The combinedorganic extracts were dried over anhydrous Na₂SO₄ and concentrated underreduced pressure. The obtained crude material was purified by silica gelcolumn chromatography (eluting with 40-45% EtOAc gradient in hexanes) toafford 37 (32 mg, 0.07 mmol, 38%) as off-white solid. ¹H NMR (500 MHz,CDCl₃): δ 9.08 (s, 1H), 8.77 (s, 1H), 8.04 (s, 1H), 7.97 (s, 1H), 7.85(d, J=8.5 Hz, 1H), 7.77-7.75 (m, 2H), 7.39-7.34 (m, 1H), 6.77-6.72 (m,1H), 6.63-6.59 (m, 1H), 5.62 (d, J=14.5 Hz, 1H), 5.13 (d, J=14.5 Hz,1H). HPLC: 99.9%. MS (ESI): m/z 438 [M+H]⁺.

Example 16

1-(6-Bromoquinoxalin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(38)

To 6-bromoquinoxalin-2(1H)-one (250 mg, 1.11 mmol) was added POBr₃ (500mg, 2.61 mmol) at RT. The reaction mixture was gradually heated to 130°C. and stirred for 2 h. After completion of reaction (by TLC), thereaction mixture was cooled to 0° C., neutralized with satd NaHCO₃solution (50 mL) and extracted with EtOAc (2×50 mL). The combinedorganic extracts were washed with H₂O (50 mL) and brine (50 mL), driedover anhydrous Na₂SO₄ and concentrated under reduced pressure to obtainthe crude material. Purification by silica gel column chromatography(eluting with 10% EtOAc/hexane) afforded compound AI (160 mg, 0.55 mmol,50%) as an off-white solid. ¹H NMR (200 MHz, CDCl₃): δ 8.84 (s, 1H),8.30 (d, J=9.0 Hz, 1H), 7.96-7.82 (m, 2H).

To a stirred suspension of copper-bronze (380 mg, 2.08 mmol) in DMSO (2mL) was added ethyl 2-bromo-2,2-difluoroacetate (0.15 mL, 1.04 mmol) atRT, and the mixture was stirred for 1 h. A solution of compound AI (150mg, 0.52 mmol) in DMSO (3 mL) was added to the reaction mixture, and thestirring was continued for another 16 h at RT. After completion ofreaction (by TLC), the reaction mixture was quenched with satd NH₄Clsolution (100 mL) and extracted with EtOAc (2×150 mL). The combinedorganic extracts were washed with H₂O (100 mL) and brine (100 mL), driedover anhydrous Na₂SO₄ and concentrated under reduced pressure to obtainthe crude material. Purification by silica gel column chromatography(eluting with 20% EtOAc/hexane) afforded ester AJ (120 mg, 0.34 mmol,69%) as an off-white solid. ¹H NMR (200 MHz, CDCl₃): δ 9.22 (s, 1H),8.34 (dd, J=8.0, 1.5 Hz, 1H), 8.02-7.94 (m, 2H), 4.40 (q, J=7.0 Hz, 2H),1.36 (t, J=7.0 Hz, 3H). MS (ESI): m/z 331 [M]⁺.

To a stirred solution of 1-bromo-2,4-difluorobenzene (0.1 mL, 0.36 mmol)in Et₂O (5 mL) was added n-BuLi (1.6 M solution in hexane; 0.22 mL, 0.36mmol) at −78° C., and the mixture was stirred for 30 min under inertatmosphere. A solution of ester AJ (0.31 g, 0.94 mmol) in Et₂O (5 mL)was added to the reaction mixture at −78° C., and stirring was continuedfor another 5 min. Progress of the reaction was monitored by TLC. Thereaction was quenched with satd NH₄Cl solution (40 mL), and the reactionmixture was extracted with EtOAc (2×50 mL). The combined organicextracts were washed with H₂O (40 mL) and brine (40 mL), dried overanhydrous Na₂SO₄ and concentrated under reduced pressure to obtain thecrude material. Purification by silica gel column chromatography(eluting with 20% EtOAc/hexane) afforded ketone AK (0.1 g, 0.65 mmol,69%) as an off-white solid. ¹H NMR (200 MHz, CDCl₃): δ 9.31 (s, 1H),8.40 (s, 1H), 8.22-7.88 (m, 3H), 7.10-6.92 (m, 1H), 6.83-6.78 (m, 1H).MS (ESI): m/z 399 [M]⁺.

To a stirred solution of ketone AK (0.35 g, 0.87 mmol) in Et₂O (15 mL)was added freshly prepared diazomethane [prepared by dissolving NMU(1.27 g, 12.5 mmol) in a 1:1 mixture of 10% KOH solution (10 mL) andEt₂O (10 mL) at 0° C. followed by separation and drying of the organiclayer using KOH pellets] at 0° C. and stirred for 30 min. The reactionmixture was allowed to warm to RT, and stirring was continued foranother 4 h. Progress of the reaction was monitored by TLC. The reactionmixture was concentrated under reduced pressure to afford the crudeproduct. Purification by silica gel column chromatography (eluting with10% EtOAc/Hexane) afforded the epoxide AL (0.24 g, 0.74 mmol, 85%) as ayellow syrup. ¹H NMR (500 MHz, CDCl₃): δ 8.99 (s, 1H), 8.18 (s, 1H),8.10-8.05 (m, 1H), 7.96-7.85 (m, 1H), 7.49-7.41 (m, 1H), 6.88-6.82 (m,1H), 6.79-6.75 (m, 1H), 3.46 (d, J=5.0 Hz, 1H), 3.03 (d, J=5.0 Hz, 1H).MS (ESI): m/z 414 [M+H]⁺.

To a stirred solution of epoxide AL (140 mg, 0.34 mmol) in dry DMF (5mL) was added 1H-tetrazole (36 mg, 0.61 mmol) followed by K₂CO₃ (56 mg,0.45 mmol) at RT under inert atmosphere. The resulting reaction mixturewas gradually heated up to 65° C. and stirred for 16 h. Progress of thereaction was monitored by TLC. The reaction mixture was diluted withice-cold H₂O (40 mL) and extracted with EtOAc (2×50 mL). The combinedorganic extracts were washed with H₂O (40 mL) and brine (40 mL), driedover anhydrous Na₂SO₄ and concentrated under reduced pressure to obtainthe crude material. Purification by silica gel column chromatography(eluting with 40% EtOAc/Hexane) to afford 38 (30 mg, 0.06 mmol, 18.4%)as a colorless thick gum. ¹H NMR (500 MHz, CDCl₃): δ 9.00 (s, 1H), 8.73(s, 1H), 8.37 (s, 1H), 8.06-7.93 (m, 2H), 7.25-7.24 (m, 1H), 6.81-6.77(m, 1H), 6.67-6.65 (m, 1H), 5.81 (s, OH), 5.72 (d, J=14.5 Hz, 1H), 5.21(d, J=14.5 Hz, 1H). HPLC: 96.5%. MS (ESI): m/z 485 [M+2]⁺.

Example 17

1-(5-(4-(Difluoromethoxy)phenyl)pyrazin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(39)

To a stirred solution of 4-bromophenol (5 g, 28.90 mmol) in acetonitrile(350 mL) was added a solution of KOH (32.5 g, 580.35 mmol) in H₂O (350mL) at 0° C., and the mixture was maintained for 5 min. To this mixture,diethyl (bromodifluoromethyl)phosphonate (9.25 mL, 52.02 mmol) was addedslowly at same temperature over 15 min (exothermic reaction), and themixture was allowed to stir at RT. After being stirred for 10 h at RT,the reaction mixture was diluted with EtOAc (100 mL), and the organiclayer was separated. The organic layer was washed with H₂O (40 mL) andbrine (40 mL), dried over anhydrous Na₂SO₄ and concentrated underreduced pressure to obtain the crude material. Purification by silicagel column chromatography (eluting with 5-10% EtOAc gradient in hexanes)afforded AM (3.0 g, 13.45 mmol, 46%). ¹H NMR (500 MHz, CDCl₃): δ 7.48(d, J=8.5 Hz, 2H), 7.01 (d, J=8.5 Hz, 2H), 6.48 (t, J_(F-H)=74.0 Hz,1H).

To a stirred solution of bromophenol derivative AM (2.0 g, 8.97 mmol) in1,4-dioxane (40 mL) were added bis(pinacolato)diboron (2.28 g, 8.97mmol) and KOAc (2.64 g, 26.90 mmol) at RT under inert atmosphere, andthe mixture was degassed for 20 min by purging with argon. To thissolution, Pd(dppf)₂Cl₂ (0.33 g, 0.45 mmol) was added, and the mixturewas degassed for a further 10 min. The reaction mixture was then heatedto 80° C. and stirred for 3 h at this temperature. Progress of thereaction was monitored by TLC. The reaction mixture was then cooled toRT and diluted with EtOAc (30 mL). The resultant solution was filteredthrough a Celite® pad, and the filtrate was then concentrated in vacuo.The crude compound was purified by silica gel column chromatography(eluting with 5-10% EtOAc gradient in hexanes) to afford AN (1.72 g,6.37 mmol, 71%). ¹H NMR (500 MHz, CDCl₃): δ 7.81 (d, J=8.5 Hz, 2H), 7.09(d, J=8.5 Hz, 2H), 6.54 (t, J_(F-H)=74.0 Hz, 1H), 1.34 (s, 12H). Astirring solution of 2,5-dibromopyrazine (1.32 g, 5.55 mmol), boronateAN (1.5 g, 5.55 mmol) and K₂CO₃ (2.27 g, 16.45 mmol) in THF-H₂O (4:1; 25mL) at RT was degassed by purging with argon for 20 min. To thissolution, Pd(dppf)₂Cl₂ (0.4 g, 0.55 mmol) was added, and the mixture wasdegassed further for 10 min. The resultant reaction mixture wasmaintained at the same temperature for 18 h; progress of the reactionwas monitored by TLC. The reaction mixture was diluted with EtOAc (30mL). The organic layer was separated and the aqueous layer was extractedwith EtOAc (2×20 mL). The combined organic layers were washed with brine(30 mL), dried over anhydrous Na₂SO₄ and concentrated under reducedpressure to obtain the crude compound. Purification by silica gel columnchromatography (eluting with 5-10% EtOAc gradient in hexanes) affordedAO (0.9 g, 2.99 mmol, 54%). ¹H NMR (500 MHz, CDCl₃): δ 8.75 (s, 1H),8.71 (s, 1H), 8.01 (d, J=9.0 Hz, 2H), 7.28 (d, J=9.0 Hz, 2H), 6.58 (t,J_(F-H)=74.0 Hz, 1H).

To a stirred suspension of copper powder (770 mg, 12.12 mmol) in DMSO (8mL) was added ethyl 2-bromo-2,2-difluoroacetate (0.77 mL, 6.0 mmol) atRT, and the mixture was stirred for 1 h. A solution of compound AO (900mg, 2.99 mmol) in DMSO (2 mL) was added to the reaction mixture andstirring was continued for another 18 h at RT. After completion ofreaction (by TLC), the reaction mixture was quenched with satd NH₄Clsolution and filtered through a Celite® pad. The Celite® pad was washedwith CH₂Cl₂ (3×50 mL). The separated organic layer was washed withbrine, dried over anhydrous Na₂SO₄ and concentrated under reducedpressure. The crude material was purified by silica gel columnchromatography (eluting with 8% EtOAc in hexanes) to afford ester AP(600 mg, 1.74 mmol, 58%). ¹H NMR (500 MHz, CDCl₃): δ 9.02 (s, 1H), 9.01(s, 1H), 8.09 (d, J=9.0 Hz, 2H), 7.28 (d, J=9.0 Hz, 2H), 6.60 (t,J_(F-H)=73.0 Hz, 1H), 4.40 (q, J=7.0 Hz, 2H), 1.35 (t, J=7.0 Hz, 3H). MS(ESI): m/z 345 [M+H]⁺.

To a stirred solution of 1-bromo-2,4-difluorobenzene (196 mg, 1.01 mmol)in Et₂O (10 mL) was added n-BuLi (2.5 M in hexanes; 0.43 mL, 1.01 mmol)dropwise at −78° C. under inert atmosphere, and the mixture was stirredfor 40 min. A solution of ester AP (350 mg, 1.01 mmol) in THF (5 mL) wasadded to the reaction mixture at −78° C., and stirring was continued foranother 10 min. The progress of the reaction was monitored by TLC. Thereaction was quenched with satd NH₄Cl solution, and the mixture wasextracted with EtOAc (2×25 mL). The combined organic extracts werewashed with brine, dried over anhydrous Na₂SO₄ and concentrated underreduced pressure to afford ketone AQ (350 mg, crude). The crude productAQ was confirmed by ¹H-NMR analysis and was taken to the next stepwithout any further purification. ¹H NMR (500 MHz, CDCl₃): δ 9.10 (s,1H), 8.95 (s, 1H), 8.11-8.07 (m, 3H), 7.29-7.26 (m, 2H), 7.04-7.01 (m,1H), 6.87-6.85 (m, 1H), 6.60 (t, J_(F-H)=73.0 Hz, 1H). MS (ESI): m/z 413[M+H]⁺.

To a stirred solution of ketone AQ (350 mg, crude) in Et₂O (15 mL) wasadded freshly prepared diazomethane [prepared by using dissolving NMU(438 mg, 4.25 mmol) in a mixture of 10% aq KOH solution (50 mL) and Et₂O(25 mL) at 0° C. followed by separation and drying of the organic layerusing KOH pellets] at 0° C., and the mixture was stirred for 3 h. Theresulting reaction mixture was allowed to stir at RT for another 30 min.The progress of the reaction was monitored by TLC. The reaction mixturewas concentrated under reduced pressure to afford the crude product.Purification by silica gel column chromatography (eluting with 15-20%EtOAc gradient in hexanes) afforded the epoxide AR (160 mg, 0.37 mmol,37% from ester AP in two steps). ¹H NMR (500 MHz, CDCl₃): δ 9.02 (s,1H), 8.75 (s, 1H), 8.09 (d, J=9.0 Hz, 2H), 7.45-7.42 (m, 1H), 7.28 (d,J=9.0 Hz, 2H), 6.89-6.83 (m, 1H), 6.78-6.75 (m, 1H), 6.60 (t,J_(F-H)=73.0 Hz, 1H), 3.47 (d, J=5.0 Hz, 1H), 3.03 (d, J=5.0 Hz, 1H).

To a stirred solution of epoxide AR (160 mg, 0.37 mmol) in dry DMF (5mL) was added 1H-tetrazole (40 mg, 0.57 mmol) followed by K₂CO₃ (52 mg,0.37 mmol) at RT under an inert atmosphere. The resulting reactionmixture was gradually heated up to 65° C. and stirred for 18 h. Theprogress of the reaction was monitored by TLC. The reaction mixture wasdiluted with ice-cold H₂O and extracted with EtOAc (2×30 mL). Thecombined organic extracts were dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure. The obtained crude material waspurified by silica gel column chromatography (eluting with 45-50% EtOAcgradient in hexanes) to afford 39 (60 mg, 0.12 mmol, 32%) as a whitesolid. ¹H NMR (500 MHz, CDCl₃): δ 8.89 (s, 1H), 8.81 (s, 1H), 8.72 (s,1H), 8.07 (d, J=9.0 Hz, 2H), 7.35-7.31 (m, 1H), 7.28 (d, J=9.0 Hz, 2H),6.81-6.77 (m, 1H), 6.75-6.69 (m, 1H), 6.60 (t, J_(F,H)=73.0 Hz, 1H),6.01 (s, OH), 5.64 (d, J=15.0 Hz, 1H), 5.18 (d, J=15.0 Hz, 1H). HPLC:95.07%. MS (ESI): m/z 497 [M+H]⁺.

Compounds 40-48 in Table 1 were prepared using the same conditions ascompound 39 from commercially available starting materials or preparedintermediates (given in Table 1).

Example 18

Chiral Preparative HPLC Separation of Enantiomers of 42

The enantiomers of 42 (300 mg, 0.58 mmol) were separated by preparativeHPLC using a CHIRALPAK IC® column (250×20 mm, 5μ) with mobile phase (A)n-hexane—(B) EtOH (A:B=90:10) and flow rate 15 mL/min to obtain 42(+)(90 mg, 0.17 mmol, 30%) as a white solid.

Analytical Data:

Chiral HPLC: 100% ee, R_(t)=15.22 min (CHIRALPAK IC® column, 250×4.6 mm,5μ; mobile phase (A) n-hexane—(B) EtOH (A:B=90:10); flow rate: 1.00mL/min). Optical rotation [α]_(D) ²⁵: +33.04° (c=0.1% in CH₃OH). ¹H NMR(500 MHz, CDCl₃): δ 8.90 (s, 1H), 8.82 (s, 1H), 8.72 (s, 1H), 8.09 (d,J=9.0 Hz, 2H), 7.38 (d, J=9.0 Hz, 2H), 7.35-7.31 (m, 1H), 6.82-6.77 (m,1H), 6.73-6.69 (m, 1H), 5.97 (s, OH), 5.64 (d, J=15.0 Hz, 1H), 5.20 (d,J=15.0 Hz, 1H). HPLC: 99.78%. MS (ESI): m/z 515 [M+H]⁺.

Example 19

1-(5-Chloropyrazin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(49)

To a stirred suspension of copper powder (3.0 g, 46.51 mmol) in DMSO (10mL) was added ethyl 2-bromo-2,2-difluoroacetate (4.7 g, 23.15 mmol) atRT, and the mixture was stirred at RT for 1 h under inert atmosphere. Asolution of 2-bromo-5-chloropyrazine (3.0 g, 15.54 mmol) in DMSO (20 mL)was added to the reaction mixture, and stirring was continued foranother 16 h at RT. After the consumption of starting material (by TLC),the reaction mixture was diluted with aq NH₄Cl solution (40 mL),filtered through a Celite® pad and washed with CH₂Cl₂ (3×25 mL). Thecollected filtrate was washed with H₂O (30 mL) and brine (30 mL), driedover anhydrous Na₂SO₄ and concentrated under reduced pressure to obtainthe crude material. Purification by column chromatography (eluting with20% EtOAc/hexane) afforded ester AS (1.12 g, 4.73 mmol, 31%) as aliquid. ¹H NMR (500 MHz, CDCl₃): δ 8.78 (s, 1H), 8.62 (s, 1H), 4.38 (q,J=7.0 Hz, 2H), 1.37 (t, J=7.0 Hz, 3H).

To a stirred solution of 1-bromo-2,4-difluorobenzene (0.98 g, 5.08 mmol)in Et₂O (20 mL) was added n-BuLi (1.6 M in hexane; 3.2 mL, 5.08 mmol)dropwise at −78° C., and the mixture was stirred for 30 min A solutionof compound AS (0.6 g, 2.54 mmol) in Et₂O (8 mL) was added to thereaction mixture at −78° C., and stirring was continued for another 5min. After consumption of the starting material (by TLC), the reactionmixture was quenched with satd NH₄Cl solution (20 mL) and extracted withEtOAc (2×50 mL). The combined organic extracts were washed with H₂O (40mL) and brine (40 mL), dried over anhydrous Na₂SO₄ and concentratedunder reduced pressure to afford ketone AT (0.7 g, crude), which wastaken on to the next reaction without further purification. ¹H NMR (500MHz, CDCl₃): δ 8.87 (s, 1H), 8.57 (s, 1H), 8.07-8.02 (m, 1H), 7.06-7.02(m, 1H), 6.90-6.84 (m, 1H).

To a stirred solution of ketone AT (0.6 g, crude) in Et₂O (10 mL) wasadded freshly prepared diazomethane [prepared by dissolving NMU (1.0 g,9.70 mmol) in 10% aq KOH (50 mL) and Et₂O (50 mL) at 0° C. followed byseparation and drying of the organic layer using KOH pellets] at 0° C.,and stirring was continued for 30 min at 0° C. The resulting reactionmixture was allowed to warm to RT and stirred for 16 h. The progress ofthe reaction was monitored by TLC. The reaction mixture was concentratedunder reduced pressure. The crude material was purified by silica gelcolumn chromatography (eluting with 20% EtOAc/hexane) to afford theepoxide AU (0.27 g, 0.84 mmol, 33% over two steps from compound AS) as asemi solid. ¹H NMR (500 MHz, CDCl₃): δ 8.62 (s, 1H), 8.51 (s, 1H),7.42-7.37 (m, 1H), 6.89-6.86 (m, 1H), 6.79-6.77 (m, 1H), 3.43 (d, J=5.0Hz, 1H), 3.00 (d, J=5.0 Hz, 1H).

To a stirred solution of epoxide AU (200 mg, 0.62 mmol) in dry DMF (6mL) were added 1H-tetrazole (65 mg, 0.93 mmol) and K₂CO₃ (86 mg, 0.62mmol) at RT under inert atmosphere. The resulting reaction mixture wasgradually heated up to 70° C. and stirred for 16 h. After consumption ofthe starting material (by TLC), the reaction mixture was cooled to RT,diluted with H₂O (30 mL) and extracted with EtOAc (2×25 mL). Thecombined organic extracts were washed with H₂O (30 mL) and brine (30mL), dried over anhydrous Na₂SO₄ and concentrated under reduced pressureto get crude mixture. The crude material was purified by silica gelcolumn chromatography (eluting with 45% EtOAc/hexane) to afford 49 (15mg, 0.038 mmol, 6.2%) as an off-white solid. ¹H NMR (500 MHz, CDCl₃): δ8.65 (s, 1H), 8.59 (s, 1H), 8.56 (s, 1H), 7.28-7.25 (m, 1H), 6.81-6.74(m, 2H), 5.63 (d, J=14.0 Hz, 1H), 5.23 (s, OH), 5.15 (d, J=14.0 Hz, 1H).HPLC: 95.27%. MS (ESI): m/z 390[M+2]⁺.

Example 20

2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-((4-fluorophenyl)ethynyl)pyrazin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(50) and2-(2,4-difluorophenyl)-1,1-difluoro-1-(5-((4-fluorophenyl)ethynyl)pyrazin-2-yl)-3-(2H-tetrazol-2-yl)propan-2-ol(51)

To a stirred solution of epoxide AU (94 mg, 0.29 mmol),1-ethynyl-4-fluorobenzene (57 mg, 0.47 mmol), Et₃N (0.1 mL, 0.72 mmol)in THF (15 mL) was added copper(I) iodide (CuI; 3 mg, 0.015 mmol) at RT.After purging with inert gas for 10 min,dichlorobis(triphenylphosphine)palladium(II) (Pd (PPh₃)₂Cl₂; 10.4 mg,0.15 mmol) was added to the reaction mixture under inert atmosphere. Thereaction mixture was gradually heated up to 70° C. and stirred for 3 h.After consumption of the starting material (by TLC), the reactionmixture was filtered through a pad of Celite® and washed with EtOAc(4×15 mL). The filtrate was washed with H₂O (50 mL), dried overanhydrous Na₂SO₄ and concentrated under reduced pressure. The crudematerial was purified by silica gel column chromatography (eluting with20% EtOAc/Hexane) to afford compound AV (40 mg, 0.099 mmol, 33%) as apale-yellow solid. ¹H NMR (500 MHz, CDCl₃): δ 8.74 (s, 1H), 8.69 (s,1H), 7.63 (dd, J=9.0, 5.5 Hz, 2H), 7.42-7.38 (m, 1H), 7.12-7.09 (m, 2H),6.89-6.86 (m, 1H), 6.79-6.75 (m, 1H), 3.46 (d, J=4.5 Hz, 1H), 3.01 (d,J=4.5 Hz, 1H).

To a stirred solution of epoxide AV (260 mg, 0.65 mmol) in dry DMF (10mL) were added sequentially K₂CO₃ (90 mg, 0.65 mmol) and 1H-tetrazole(70 mg, 0.97 mmol) at RT under an inert atmosphere. The reaction mixturewas gradually heated up to 65° C. and stirred for 16 h. Afterconsumption of the starting material (by TLC), the reaction mixture wascooled to RT, diluted with H₂O (50 mL) and extracted with EtOAc (2×50mL). The separated organic layer was dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure to obtain the crude material.Purification by silica gel column chromatography afforded 51 (15 mg,0.03 mmol, 4.6%) (eluting with 25% EtOAc/hexane) as an off-white solidand 50 (30 mg, 0.06 mmol, 9.2%) (eluting with 35% EtOAc/hexane) as alight brown solid. 50: ¹H NMR (500 MHz, CDCl₃): δ 8.74 (s, 1H), 8.71 (s,1H), 8.62 (s, 1H), 7.63-7.60 (dd, J=13.5, 7.5 Hz, 2H), 7.30-7.28 (m,1H), 7.13-7.09 (m, 2H), 6.81-6.77 (m, 1H), 6.74-6.70 (m, 1H), 5.64 (d,J=14.5 Hz, 1H), 5.61 (s, OH), 5.17 (d, J=14.5 Hz, 1H). HPLC: 93.5%. MS(ESI): m/z 472 [M−H]⁻. 51: ¹H NMR (500 MHz, CDCl₃): δ 8.72 (s, 1H), 8.62(s, 1H), 8.38 (s, 1H), 7.64-7.62 (m, 2H), 7.30-7.27 (m, 1H), 7.12-7.09(m, 2H), 6.81-6.77 (m, 1H), 6.72-6.69 (m, 1H), 6.02 (d, J=14.5 Hz, 1H),5.37 (d, J=14.5 Hz, 1H), 5.10 (s, OH). HPLC: 98.3%. MS (ESI): m/z 472[M−H]⁻.

Example 21

2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(4-fluorophenethyl)pyrazin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(52)

To a stirred solution of 50 (25 mg, 0.053 mmol) in EtOAc (10 mL) wasadded 10% palladium on carbon (Pd/C; 5 mg) under inert atmosphere, andthe mixture was stirred at RT for 2 h under H₂ atmosphere (balloonpressure). After consumption of the starting material (by TLC), thereaction mixture was filtered through a pad of Celite® and washed withEtOAc (3×10 mL). The filtrate was concentrated under reduced pressure toobtain the crude material. Purification by silica gel columnchromatography (eluting with 30% EtOAc/hexane) afforded 52 (22 mg, 0.046mmol, 88%) as a brown semi-solid. ¹H NMR (500 MHz, CDCl₃): δ 8.72 (s,1H), 8.70 (s, 1H), 8.18 (s, 1H), 7.28-7.24 (m, 1H), 7.07-7.05 (m, 2H),6.96-6.93 (m, 2H), 6.80-6.76 (m, 1H), 6.70-6.67 (m, 1H), 6.08 (s, OH),5.59 (d, J=14.5 Hz, 1H), 5.15 (d, J=14.5 Hz, 1H), 3.15 (t, J=7.0 Hz,2H), 3.05 (t, J=7.0 Hz, 2H). HPLC: 88.8%. MS (ESI): m/z 477 [M+H]⁺.

Example 22

2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(4-fluorophenethyl)pyrazin-2-yl)-3-(2H-tetrazol-2-yl)propan-2-ol(53)

To a stirred solution of 51 (30 mg, 0.063 mmol) in EtOAc (10 mL) wasadded 10% Pd/C (6 mg) under inert atmosphere, and the mixture wasstirred at RT for 2 h under H₂ atmosphere (balloon pressure). Afterconsumption of the starting material (by TLC), the reaction mixture wasfiltered through a pad of Celite® and washed with EtOAc (3×10 mL). Thefiltrate was concentrated under reduced pressure to afford 53 (23 mg,0.05 mmol, 76%) as a brown semi-solid. ¹H NMR (500 MHz, CDCl₃): δ 8.69(s, 1H), 8.34 (s, 1H), 8.22 (s, 1H), 7.29-7.27 (m, 1H), 7.09-7.07 (m,2H), 6.97-6.93 (m, 2H), 6.80-6.76 (m, 1H), 6.69-6.66 (m, 1H), 5.95 (d,J=14.5 Hz, 1H), 5.34 (d, J=14.5 Hz, 1H), 5.32 (s, OH), 3.15 (t, J=7.5Hz, 2H), 3.05 (t, J=7.5 Hz, 2H). HPLC: 91.7%. MS (ESI): m/z 477 [M+H]⁺.

Example 23

2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-(trifluoromethoxy)quinoxalin-2-yl)propan-2-ol(54)

To a stirred solution of 2-nitro-5-(trifluoromethoxy)aniline (5.0 g,22.5 mmol) in EtOH (50 mL) was added 10% Pd/C (1.2 g) under inertatmosphere. The resulting reaction mixture was stirred for 16 h at RTunder H₂ atmosphere. After consumption of the starting material (byTLC), the reaction mixture was filtered through a pad of Celite® andwashed with EtOAc (3×50 mL). The filtrate was concentrated under reducedpressure to afford compound AW (4.0 g, 20.83 mmol, 93%) as an orangesyrup. ¹H NMR (500 MHz, DMSO-d₆): δ 6.50-6.45 (m, 2H), 6.29 (d, J=8.0Hz, 1H), 4.76-4.75 (br s, 4H). MS (ESI): m/z 194 [M+2]⁺.

To a stirred solution of compound AW (4.0 g, 20.83 mmol) in CH₃OH (40mL) was added 2-oxoacetic acid (2.3 mL, 20.83 mmol) at 0° C. Theresulting reaction mixture was allowed to warm to RT and stirred for 24h. After the consumption of starting material (by TLC), the reactionmixture was diluted with H₂O (50 mL) and stirred for 5 min. The solidprecipitated was filtered and washed with H₂O (3×50 mL). The crude solidwas purified by silica gel column chromatography (eluting with 30%EtOAc/hexane) to afford compound AX (1.4 g, 6.08 mmol, 29.8%) aspale-yellow solid. ¹H NMR (500 MHz, DMSO-d₆): δ 12.6 (br s, 1H), 8.25(s, 1H), 7.78 (s, 1H), 7.60 (d, J=7.0 Hz, 1H), 7.04 (d, J=8.5 Hz, 1H).MS (ESI): m/z 230 [M+H]⁺.

To compound AX (0.85 g, 3.69 mmol) was added POBr₃ (2.1 g, 7.34 mmol) atRT. The reaction mixture was gradually heated to 130° C. and stirred for2 h. After complete consumption of the starting material (by TLC), thereaction mixture was cooled to RT, diluted with ice cold H₂O (30 mL),made basic (pH ˜8) using satd NaHCO₃ solution (25 mL) and extracted withEtOAc (2×50 mL). The combined organic extracts were washed with H₂O (50mL) and brine (50 mL), dried over anhydrous Na₂SO₄ and concentratedunder reduced pressure to obtain the crude material. Purification bysilica gel column chromatography (eluting with 2% EtOAc/hexane) toafford compound AY (0.65 g, 2.22 mmol, 65%) as an off-white solid. ¹HNMR (500 MHz, CDCl₃): δ 8.89 (s, 1H), 8.09 (d, J=9.0 Hz, 1H), 7.94 (s,1H), 7.65 (dd, J=9.0, 2.0 Hz, 1H). MS (ESI): m/z 295.9 [M+2]⁺.

To a stirred suspension of copper powder (0.56 g, 0.89 mmol) in DMSO (10mL) was added ethyl 2-bromo-2,2-difluoroacetate (0.9 g, 4.45 mmol), andthe mixture was stirred at RT for 1 h. To the resulting reaction mixturewas added compound AY (0.65 g, 2.22 mmol), and stirring was continuedfor 16 h at RT. After complete consumption of the starting material (byTLC), the reaction mixture was quenched with satd NH₄Cl solution (100mL) and extracted with EtOAc (3×30 mL). The combined organic extractswere washed with H₂O (50 mL) and brine (50 mL), dried over anhydrousNa₂SO₄ and concentrated under reduced pressure to obtain the crudematerial. Purification by silica gel column chromatography (eluting with2% EtOAc/hexane) to afford ester AZ (0.55 g, 1.63 mmol, 73.6%) as apale-yellow syrup. ¹H NMR (500 MHz, CDCl₃): δ 9.26 (s, 1H), 8.21 (d,J=9.5 Hz, 1H), 8.02 (s, 1H), 7.71 (d, J=9.5 Hz, 1H), 4.42 (q, J=7.0 Hz,2H), 1.36 (t, J=7.0 Hz, 3H).

To a stirred solution of 1-bromo-2,4-difluorobenzene (0.27 mL, 2.45mmol) in Et₂O (5 mL) was added n-BuLi (1.6 M in hexane; 1.5 mL, 2.45mmol) at −78° C., and the mixture was stirred for 30 min under inertatmosphere. A solution of ester AZ (0.55 g, 1.63 mmol) in THF (5 mL) wasadded to the reaction mixture at −78° C., and stirring was continued foranother 1 h. After complete consumption of the starting material (byTLC), the reaction mixture was quenched with satd NH₄Cl solution (50 mL)and extracted with EtOAc (2×25 mL). The combined organic extracts werewashed with H₂O (25 mL) and brine (25 mL), dried over anhydrous Na₂SO₄and concentrated under reduced pressure to obtain the crude product.Purification by silica gel column chromatography (eluting with 5%EtOAc/hexane) to afford ketone BA (0.4 g, 0.98 mmol, 60%) as a yellowsyrup. ¹H NMR (500 MHz, CDCl₃): δ 9.34 (s, 1H), 8.12-8.11 (m, 2H), 8.03(s, 1H), 7.67-7.65 (m, 1H), 7.07-7.05 (m, 1H), 6.83-6.79 (m, 1H). MS(ESI): 405 [M+H]⁺.

To a stirred solution of ketone BA (0.4 g, 0.99 mmol) in Et₂O (15 mL)was added freshly prepared diazomethane [prepared by dissolving NMU (509mg, 4.95 mmol) in a 1:1 mixture of 10% KOH solution (40 mL) and Et₂O (40mL) at 0° C. followed by separation and drying of the organic layerusing KOH pellets] at 0° C. and stirred for 30 min. The resultingreaction mixture was allowed to warm to RT, and stirring was continuedfor another 4 h. After complete consumption of the starting material (byTLC), the volatiles were evaporated under reduced pressure. The crudematerial was purified by silica gel column chromatography (eluting with2% EtOAc/hexane) to afford epoxide BB (0.29 g, 0.69 mmol, 70%) as ayellow syrup. ¹H NMR (500 MHz, CDCl₃): δ 9.02 (s, 1H), 8.22 (d, J=9.5Hz, 1H), 8.00 (s, 1H), 7.70 (dd, J=9.5, 2.5 Hz, 1H), 7.46-7.43 (m, 1H),6.89-6.85 (m, 1H), 6.78-6.74 (m, 1H), 3.46 (d, J=5.0 Hz, 1H), 3.03 (d,J=5.0 Hz, 1H). MS (ESI): 419 [M+H]⁺.

To a stirred solution of epoxide BB (0.29 g, 0.69 mmol) in dry DMF (5mL) was added the sodium salt of tetrazole (95 mg, 1.03 mmol) at RTunder inert atmosphere. The resulting reaction mixture was graduallyheated up to 65° C. and stirred for 16 h. After complete consumption ofthe starting material (by TLC), the reaction mixture was diluted withice cold H₂O (30 mL) and extracted with EtOAc (2×25 mL). The combinedorganic extracts were dried over anhydrous Na₂SO₄ and concentrated underreduced pressure. The obtained crude material was purified by silica gelcolumn chromatography (eluting with 30% EtOAc/hexane) to afford 54 (140mg, 0.28 mmol, 41.4%) as an off-white solid. ¹H NMR (500 MHz, CDCl₃): δ9.04 (s, 1H), 8.72 (s, 1H), 8.13 (d, J=9.5 Hz, 1H), 8.00 (s, 1H), 7.73(dd, J=9.5, 2.0 Hz, 1H), 7.30-7.28 (m, 1H), 6.80-6.78 (m, 1H), 6.69-6.68(m, 1H), 5.73 (d, J=14.5 Hz, 1H), 5.67 (s, OH), 5.21 (d, J=14.5 Hz, 1H).HPLC: 98.3%. MS (ESI): m/z 489 [M+H]⁺.

Compound 55 in Table 1 was prepared using the same conditions ascompound 54 from commercially available starting materials or preparedintermediates (given in Table 1).

Example 24

2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-(4-(trifluoromethyl)phenyl)pyridazin-3-yl)propan-2-ol(56)

To a stirred solution of 3,6-dibromopyridazine (200 mg, 0.84 mmol) and4-(trifluoromethyl)phenylboronic acid (159.7 mg, 0.84 mmol) in1,2-dimethoxyethane (DME; 12 mL) was added 1M sodium carbonate (Na₂CO₃;1.2 mL, 1.26 mmol) at RT, and the mixture was degassed by purging withargon for 30 min. To the resulting reaction mixture was addedtetrakis(triphenylphosphine)palladium(0) (Pd(PPh₃)₄; 29.1 mg, 0.025mmol), and the mixture was further degassed for 5 min at RT. Thereaction mixture was stirred at reflux for 18 h. After completeconsumption of the starting material (by TLC), the reaction mixture wascooled to RT, diluted with H₂O (50 mL) and extracted with EtOAc (2×50mL). The combined organic extracts were washed with H₂O (40 mL) andbrine (40 mL), dried over anhydrous Na₂SO₄ and concentrated underreduced pressure to obtain the crude material. Purification by silicagel column chromatography (eluting with 12% EtOAc/hexane) afforded amixture of mono and bis-coupled products BC (150 mg, 2:1 ratio), whichwas taken to the next step without separation. (Note: Both compoundseluted at same R_(f); all the characteristic protons were seen in the ¹HNMR spectrum.) MS (ESI): 303 [M+H]⁺.

To a stirred suspension of copper powder (0.75 g, 11.81 mmol) in DMSO (3mL) was added ethyl 2-bromo-2,2-difluoroacetate (1.2 g, 5.92 mmol) atRT, and the mixture was stirred for 1 h. A solution of compound BC (0.9g, mixture) in DMSO (7 mL) was added to the reaction mixture, andstirring was continued for another 18 h at RT. After completion ofreaction (by TLC), the reaction mixture was quenched with satd NH₄Clsolution (100 mL) and extracted with EtOAc (2×200 mL). The combinedorganic extracts were washed with H₂O (50 mL) and brine (50 mL), driedover anhydrous Na₂SO₄ and concentrated under reduced pressure to obtainthe crude product. Purification by silica gel column chromatography(eluting with 15% EtOAc/hexane) afforded crude BD (0.7 g, as a mixture)which was taken for the next step without separation. (Note: All thecharacteristic protons were seen in the ¹H NMR spectrum.) LC-MS: 347.8[M+H]⁺ at 4.99 RT (73.75% purity).

To a stirred solution of 1-bromo-2,4-difluorobenzene (83.67 mg, 0.43mmol) in THF (5 mL) was added n-BuLi (1.6 M in hexane; 0.27 mL, 0.43mmol) at −78° C. and stirred for 1 h under inert atmosphere. A solutionof ester BD (100 mg, crude) in THF (3 mL) was added to the reactionmixture at −78° C., and stirring was continued for another 2 h. Theprogress of the reaction was monitored by TLC. The reaction was quenchedwith satd NH₄Cl solution (50 mL) and extracted with EtOAc (2×50 mL). Thecombined organic extracts were washed with H₂O (50 mL) and brine (50mL), dried over anhydrous Na₂SO₄ and concentrated under reduced pressureto afford ketone BE (100 mg, crude). The mixture was taken on to thenext step without purification.

(Note: All the characteristic protons were seen in ¹H NMR spectrum.) MS(ESI): 415 [M+H]⁺. To a stirred solution of ketone BE (100 mg, crude) inEt₂O (20 mL) was added freshly prepared diazomethane [prepared bydissolving NMU (240 mg, 2.41 mmol) in a 1:1 mixture of 10% KOH solution(40 mL) and Et₂O (40 mL) at 0° C. followed by separation and drying ofthe organic layer using KOH pellets] at 0° C., and the mixture wasstirred for 1 h. The resulting reaction mixture was allowed to warm toRT, and stirring was continued for another 5 h. The progress of thereaction was monitored by TLC. The reaction mixture was concentratedunder reduced pressure to afford BF (60 mg, crude). The obtained crudemixture was taken to the next step without purification. (Note: All thecharacteristic protons were seen in the ¹H NMR spectrum.) MS (ESI):429.9 [M+H]⁺.

To a stirred solution of epoxide BF (60 mg, crude) in dry DMF (4 mL) wasadded 1H-tetrazole (19.25 mg, 0.27 mmol) followed by K₂CO₃ (19.25 mg,0.14 mmol) at RT under an inert atmosphere. The resulting reactionmixture was gradually heated up to 65° C. and stirred for 20 h. Theprogress of the reaction was monitored by TLC. The reaction mixture wasdiluted with ice cold H₂O (50 mL) and extracted with EtOAc (2×50 mL).The combined organic extracts were washed with H₂O (50 mL) and brine (50mL), dried over anhydrous Na₂SO₄ and concentrated under reducedpressure. The obtained crude material was purified by preparative TLC(eluting with 40% EtOAc/hexane; R_(f)=0.2) to afford 56 (11.5 mg, 0.02mmol) as a colorless semi-solid. ¹H NMR (500 MHz, CDCl₃): δ 8.80 (s,1H), 8.19-8.17 (m, 2H), 8.01 (d, J=8.5 Hz, 1H), 7.83-7.81 (m, 3H),7.41-7.37 (m, 1H), 6.80-6.68 (m, 3H), 5.70 (d, J=14.5 Hz, 1H), 5.28 (d,J=14.5 Hz, 1H). HPLC: 97.6%. MS (ESI): m/z 499.4 [M+H]⁺.

Compounds 57 and 58 in Table 1 were prepared using the same conditionsas compound 56 from commercially available starting materials (given inTable 1).

Example 25

2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-vinylquinoxalin-2-yl)propan-2-ol(59)

To 6-bromoquinoxalin-2(1H)-one (1.0 g, 4.44 mmol) was added POBr₃ (2.54g, 8.88 mmol) at RT. The reaction mixture was gradually heated to 130°C. and stirred for 2 h. After complete consumption of the startingmaterial (by TLC), the reaction mixture was cooled to RT, neutralizedwith satd NaHCO₃ solution at 0° C. and extracted with EtOAc (2×100 mL).The combined organic extracts were washed with H₂O (100 mL) and brine(100 mL), dried over anhydrous Na₂SO₄ and concentrated under reducedpressure to obtain the crude product. Purification by silica gel columnchromatography (eluting with 10% EtOAc/hexane) afforded compound BG (2.5g, 9.0 mmol, 40%) as an off-white solid. ¹H NMR (500 MHz, CDCl₃): δ 8.84(s, 1H), 8.30 (d, J=9.0 Hz, 1H), 7.96-7.82 (m, 2H).

To a stirred suspension of copper powder (0.77 g, 12.1 mmol) in DMSO (15mL) was added ethyl 2-bromo-2,2-difluoroacetate (1.23 g, 6.09 mmol) atRT, and the reaction mixture was stirred for 1 h. A solution of compoundBG (0.85 g, 3.04 mmol) in DMSO (5 mL) was added to the reaction mixture,and stirring was continued for another 16 h at RT. After completeconsumption of the starting material (by TLC), the reaction mixture wasquenched with satd NH₄Cl solution (50 mL) and extracted with EtOAc (2×25mL). The combined organic extracts were washed with H₂O (50 mL) andbrine (50 mL), dried over anhydrous Na₂SO₄ and concentrated underreduced pressure to obtain the crude material. Purification by silicagel column chromatography (eluting with 20% EtOAc/hexane) afforded esterBH (0.53 g, 1.6 mmol, 53%) as an off-white solid. ¹H NMR (500 MHz,CDCl₃): δ 9.22 (s, 1H), 8.34 (dd, J=8.0, 1.5 Hz, 1H), 8.02-7.94 (m, 2H),4.40 (q, J=7.0 Hz, 2H), 1.36 (t, J=7.0 Hz, 3H). MS (ESI): m/z 332[M+H]⁺.

To a stirred solution of 1-bromo-2,4-difluorobenzene (0.32 g, 1.66 mmol)in Et₂O (30 mL) was added n-BuLi (1.6 M in hexane; 0.1 mL, 1.66 mmol) at−78° C., and the reaction mixture was stirred for 30 min under inertatmosphere. A solution of ester BH (0.55 g, 1.66 mmol) in Et₂O (10 mL)was added to the reaction mixture at −78° C., and stirring was continuedfor another 5 min. After complete consumption of the starting material(by TLC), the reaction mixture was quenched with satd NH₄Cl solution (50mL) and extracted with EtOAc (2×25 mL). The combined organic extractswere washed with H₂O (25 mL) and brine (25 mL), dried over anhydrousNa₂SO₄ and concentrated under reduced pressure to obtain the crudeproduct. Purification by silica gel column chromatography (eluting with20% EtOAc/hexane) afford ketone BI (0.58 g, 1.46 mmol, 88%) as anoff-white solid. ¹H NMR (500 MHz, CDCl₃): δ 9.31 (s, 1H), 8.40 (s, 1H),8.22-7.88 (m, 3H), 7.10-6.92 (m, 1H), 6.83-6.78 (m, 1H). MS (ESI): m/z400 [M+H]⁺.

To a stirred solution of ketone BI (0.59 g, 1.46 mmol) in Et₂O (10 mL)was added freshly prepared diazomethane [prepared by dissolving NMU(0.75 g, 7.3 mmol) in a 1:1 mixture of 10% KOH solution (20 mL) and Et₂O(20 mL) at 0° C. followed by separation and drying of the organic layerusing KOH pellets] at 0° C., and the mixture was stirred for 30 min. Theresulting reaction mixture was allowed to warm to RT, and stirring wascontinued for another 4 h. After complete consumption of the startingmaterial (by TLC), the reaction mixture was concentrated under reducedpressure to afford the crude product. Purification by silica gel columnchromatography (eluting with 10% EtOAc/hexane) afforded the epoxide BJ(0.53 g, 1.28 mmol, 88%) as a yellow syrup. ¹H NMR (500 MHz, CDCl₃): δ8.99 (s, 1H), 8.18 (s, 1H), 8.10-8.05 (m, 1H), 7.96-7.85 (m, 1H),7.49-7.41 (m, 1H), 6.88-6.82 (m, 1H), 6.79-6.75 (m, 1H), 3.46 (d, J=5.0Hz, 1H), 3.03 (d, J=5.0 Hz, 1H). MS (ESI): m/z 414 [M+H]⁺.

A stirred solution of compound BJ (0.32 g, 0.72 mmol) and tetravinyltin(0.115 g, 0.72 mmol) in 1,4-dioxane (20 mL) was degassed by purging withinert gas for 10 min at RT. To the reaction mixture was added Pd(PPh₃)₄(0.08 g, 0.073 mmol), and the mixture was degassed for another 10 min atRT. The reaction mixture was then stirred for 3 h at 70° C. Aftercomplete consumption of the starting material (by TLC), the reactionmixture was cooled to RT, filtered through a pad of Celite® and thefiltrate was concentrated under reduced pressure to obtain the crudematerial. Purification by silica gel column chromatography (eluting with7% EtOAc/hexane) afforded compound BK (0.15 g, 0.41 mmol, 57%) as acolorless liquid. This material contained some tin impurities and wasused in the next step without further purification. ¹H NMR (500 MHz,CDCl₃): δ 8.95 (s, 1H), 8.11-8.09 (m, 2H), 8.10 (d, J=9.5 Hz, 1H),7.99-7.97 (m, 1H), 7.45-7.43 (m, 1H), 6.87-6.84 (m, 1H), 6.77-6.74 (m,1H), 6.04 (d, J=17.5 Hz, 1H), 5.55 (d, J=11.5 Hz, 1H), 3.46 (d, J=5.0Hz, 1H), 3.03 (d, J=5.0 Hz, 1H). MS (ESI): m/z 361 [M+H]⁺.

To a stirred solution of epoxide BK (150 mg, 0.41 mmol) in dry DMF (10mL) was added 1H-tetrazole (43 mg, 0.61 mmol) followed by K₂CO₃ (57 mg,0.41 mmol) at RT under an inert atmosphere. The resulting reactionmixture was gradually heated up to 65° C. and stirred for 16 h. Aftercomplete consumption of the starting material (by TLC), the reactionmixture was diluted with ice-cold H₂O (30 mL) and extracted with EtOAc(2×30 mL). The combined organic extracts were washed with H₂O (30 mL)and brine (30 mL), dried over anhydrous Na₂SO₄ and concentrated underreduced pressure to obtain the crude product. Purification by silica gelcolumn chromatography (eluting with 40% EtOAc/hexane) afforded 59 (35mg, 0.08 mmol, 20%) as a colorless thick syrup. ¹H NMR (500 MHz, CDCl₃):δ 8.98 (s, 1H), 8.74 (s, 1H), 8.06 (s, 1H), 8.02 (d, J=9.5 Hz, 2H),7.31-7.29 (m, 1H), 6.96-6.93 (m, 1H), 6.80-6.76 (m, 1H), 6.65-6.63 (m,1H), 6.22 (s, 1H), 6.06 (d, J=17.5 Hz, 1H), 5.71 (d, J=14.5 Hz, 1H),5.60 (d, J=11 Hz, 1H) 5.21 (d, J=14.5 Hz, 1H). HPLC: 94%. MS (ESI): m/z431 [M+H]⁺.

Example 26

2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(4′-(2,2,2-trifluoroethoxy)biphenyl-4-yl)propan-2-ol (60)

To a suspension of copper powder (1.8 g, 28.3 mmol) in DMSO (20 mL) wasadded ethyl 2-bromo-2,2-difluoroacetate (1.8 mL, 14.13 mmol), and themixture was stirred for 1 h at RT under an inert atmosphere. To theresulting solution was added 1-bromo-4-iodobenzene (2.0 g, 7.07 mmol),and stirring was continued for 10 h at RT. After complete consumption ofthe starting material (by TLC), the reaction mixture was quenched withsatd NH₄Cl solution (30 mL) and extracted with CH₂Cl₂ (3×50 mL). Thecombined organic extracts were washed with H₂O (30 mL) and brine (30mL), dried over anhydrous Na₂SO₄ and concentrated under reduced pressureto obtain the crude product. Purification by silica gel columnchromatography (eluting with 2.5% EtOAc/hexane) afforded the ester BL(2.1 g, 7.53 mmol, 72%) as a liquid. ¹H NMR (500 MHz, CDCl₃): δ 7.59 (d,J=9.0 Hz, 2H), 7.48 (d, J=9.0 Hz, 2H), 4.30 (q, J=7.0 Hz, 2H), 1.30 (t,J=7.0 Hz, 3H).

To a stirred solution of 1-bromo-2,4-difluorobenzene (0.2 mL, 1.79 mmol)in Et₂O (5 mL) was added n-BuLi (1.6 M in hexanes; 1.1 mL, 1.79 mmol) at−78° C., and the mixture was stirred for 30 min under inert atmosphere.A solution of ester BL (500 mg, 1.79 mmol) in Et₂O (5 mL) was added tothe reaction mixture at −78° C., and stirring was continued for another2 h. After complete consumption of the starting material (by TLC), thereaction mixture was quenched with satd NH₄Cl solution and extractedwith CH₂Cl₂ (2×50 mL). The combined organic extracts were washed withH₂O (30 mL) and brine (30 mL), dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure to obtain the crude product.Purification by silica gel column chromatography (eluting with 3%EtOAc/hexane) afforded the ketone BM (400 mg, 1.15 mmol, 64%) as apale-yellow liquid. ¹H NMR (500 MHz, CDCl₃): δ 7.85-7.80 (m, 1H), 7.61(d, J=8.5 Hz, 2H), 7.47 (d, J=8.5 Hz, 2H), 6.99-6.96 (m, 1H), 6.89-6.85(m, 1H).

To a stirred solution of ketone BM (1.7 g, 4.91 mmol) in Et₂O (15 mL)was added freshly prepared diazomethane [prepared by dissolving NMU (2.5g, 24.56 mmol) in a 1:1 mixture of 10% KOH solution (25 mL) and Et₂O (25mL) at 0° C. followed by separation and drying of the organic layerusing KOH pellets] at −5° C., and the mixture was stirred for 2 h. Theresulting reaction mixture was allowed to warm to RT, and stirring wascontinued for another 16 h. Progress of the reaction was monitored byTLC. The reaction mixture was then concentrated under reduced pressureto obtain the crude product. Purification by silica gel columnchromatography (eluting with 2-3% EtOAc/hexane) afforded the epoxide BN(1.5 g, 4.9 mmol, 88%) as a semi-solid. ¹H NMR (200 MHz, CDCl₃): δ7.55-7.51 (m, 2H), 7.29-7.21 (m, 3H), 7.86-6.71 (m, 2H), 3.25 (d, J=5.0Hz, 1H), 2.97-2.91 (m, 1H).

To a stirred solution of epoxide BN (100 mg, 0.33 mmol) in THF-H₂O (15mL, 8:2 v/v) were added sequentially Na₂CO₃ (90 mg, 0.83 mmol) andboronate I-F (120 mg, 0.33 mmol) at RT under an inert atmosphere. Afterpurging with nitrogen for 10 min, Pd(dppf)₂Cl₂ (68 mg, 0.083 mmol) wasadded to reaction mixture under an inert atmosphere, and the resultingmixture was stirred at 70° C. for 4 h. The reaction mixture was allowedto cool to RT, diluted with H₂O (15 mL) and extracted with EtOAc (2×50mL). The combined organic extracts were washed with H₂O (20 mL) andbrine (20 mL), dried over anhydrous Na₂SO₄ and concentrated in vacuo.The crude material was purified by silica gel column chromatography(eluting with 3-4% EtOAc/hexane) to afford BO (110 mg, 0.26 mmol, 86%)as white solid. ¹H NMR (200 MHz, CDCl₃): δ 7.62-7.43 (m, 5H), 7.28-7.22(m, 3H), 7.04-7.00 (m, 1H), 6.84-6.76 (m, 2H), 4.40 (q, J=8.0 Hz, 2H),3.28 (dd, J=5.6, 2.0 Hz, 1H), 2.95-2.92 (m, 1H). MS (ESI): m/z 455[M−H]⁻.

To a stirred solution of epoxide BO (120 mg, 0.26 mmol) in dry DMF (5mL) was added 1H-tetrazole (28 mg, 0.39 mmol) followed by K₂CO₃ (73 mg,0.52 mmol) at RT under an inert atmosphere. The resulting reactionmixture was gradually heated up to 65° C. and stirred for 8 h; progressof the reaction was monitored by TLC. The reaction mixture was thendiluted with ice-cold H₂O and extracted with EtOAc (3×20 mL). Theseparated organic layer was washed with H₂O (50 mL) and brine (50 mL),dried over anhydrous Na₂SO₄ and concentrated under reduced pressure. Thecrude material was purified by silica gel column chromatography (elutingwith 30% EtOAc/hexane) to afford 60 (20 mg, 0.04 mmol, 15%) as apale-yellow solid. ¹H NMR (500 MHz, CDCl₃): δ 8.65 (s, 1H), 7.53 (d,J=9.0 Hz, 2H), 7.49 (d, J=7.5 Hz, 2H), 7.25 (d, J=8.0 Hz, 1H), 7.17-7.12(m, 1H), 7.07-7.01 (m, 3H), 6.78-6.75 (m, 1H), 6.68-6.66 (m, 1H), 5.70(d, J=14.5 Hz, 1H), 5.07 (d, J=14.5 Hz, 1H), 4.42-4.37 (q, J=8.0 Hz,2H). MS (ESI): m/z 527 [M+H]⁺.

Compound 61 in Table 1 was prepared using the same conditions ascompound 60 from commercially available starting materials (given inTable 1).

-   HPLC Method A Specifications-   Column: Aquity BEH C-18 (50×2.1 mm, 1.7μ)-   Mobile Phase: A) Acetonitrile; B) 0.025% aq TFA-   Flow Rate: 0.50 mL/min-   Time (min)/% B: 0.01/90, 0.5/90, 3/10, 6/10

TABLE 1 Structures for Example Compounds Compound Number StructureStarting Material  1

See Example 1  2

See Example 2  3

See Example 3  4

See Example 4  5

See Example 5  6

See Example 6  7

See Example 7  8

See Example 8  9

See Example 9 10

See Example 10 11

See Example 11 12

2-bromothiazole 13

2,5-dibromothiophene 14

2-iodothiophene 15

1-bromo-4-methoxybenzene 16

2-bromo-6-chloroquinoline; 2- bromo-6-chloroquinoline 17

6-fluoroquinoline 18

6-(trifluoromethyl)quinoline 19

2-bromo-6-(2,2,2- trifluoroethoxy)quinoline (I-A, Example 12) 20

1-bromo-2-fluoro-4- (trifluoromethyl)benzene; 2- bromo-6-chloroquinoline21

2-bromo-6- (trifluoromethoxy)quinoline (I-B, Example 12) 22

1-bromo-2-chloro-4- (trifluoromethyl)benzene; 2- bromo-6-chloroquinoline23

4-bromo-1,2-difluorobenzene; 2- bromo-6-chloroquinoline 24

quinoline-6-carbonitrile (I-C, Example 12) 25

6-(difluoromethyl)quinoline (I-D, Example 12) 26

6-methylquinoline 27

2,6-dibromobenzo[d]thiazole 28

2-bromo-1,4-difluorobenzene; 2- bromo-6-chloroquinoline 29

5,6-dichloroquinoline 30

5-(2,2,2- trifluoroethoxy)quinoline (I-E, Example 12) 31

5-chloroquinoline 32

1-bromo-4-fluorobenzene; 2- bromo-6-chloroquinoline 33

1,2,4-triazole; 2-bromo-6- chloroquinoline 34

1-bromo-4-chloro-2- fluorobenzene; 2-bromo-6- chloroquinoxaline 35

See Example 13 36

See Example 14 37

See Example 15 38

See Example 16 39

See Example 17 40

(4-chlorophenyl)boronic acid 41

4,4,5,5-tetramethyl-2-(4-(2,2,2- trifluoroethoxy)phenyl)-1,3,2-dioxaborolane (I-F, Example 12) 42

(4- (trifluoromethoxy)phenyl)boronic acid 43

(4-bromophenyl)boronic acid 44

(3,4-difluorophenyl)boronic acid 45

1-bromo-4-fluorobenzene 46

(4-fluorophenyl)boronic acid 47

(4-fluorophenyl)boronic acid 48

(4-methoxyphenyl)boronic acid 49

See Example 19 50

See Example 20 51

See Example 20 52

See Example 21 53

See Example 22 54

See Example 23 55

4-fluorobenzene-1,2-diamine 56

See Example 24 57

(4- (trifluoromethoxy)phenyl)boronic acid 58

(4-fluorophenyl)boronic acid 59

See Example 25 60

See Example 26 61

(4-fluorophenyl)boronic acid

TABLE 2 Analytical Data for Example Compounds in Table 1 HPLC RetentionPatent HPLC Time ESIMS Example# Method (min) (M + H)  1 A 2.53 393  2 A2.32 438  3 A 2.48 460.9  4 A 2.62 438  4(−) A 2.67 438  5 A 2.44 404  6A 2.44 410  7 A 1.82 355  8(−) A 2.76 454  9(+) A 2.6 484 (M + 2) 10(−)A 2.51 439 11 A 2.65 444.6 12 A 2.08 360 13 A 2.57 453 (M − 1) 14 A 2.37359 15 A 2.53 432 16 A 2.78 438 17 A 2.5 422 18 A 2.65 472 19 A 2.71 50220 A 2.82 488 21 A 2.73 488 22 A 2.84 504.8 23 A 2.62 438 24 A 2.33 42925 A 2.46 454 26 A 2.66 418.2 27 A 2.73 490.2 28 A 2.63 438 29 A 2.83472.8 30 A 2.67 502.4 31 A 2.63 438 32 A 2.56 420 33(+) A 2.57 437 34 A2.55 455 35 A 2.32 437 36 A 2.69 528 37 A 2.58 438 38 A 2.56 483 39 A2.61 497 40 A 2.78 465.1 41 A 2.72 529 42 A 2.85 515 43 A 2.85 515.443(+) A 2.81 509 44 A 2.62 467.4 45 A 2.77 497.4 46 A 2.52 447.4 (M − 1)47 A 2.65 447.3 (M − 1) 48 A 2.52 461.3 49 A 2.21 387.4 (M − 1) 50 A2.65 473.9 51 A 2.77 473.8 52 A 2.57 477.6 53 A 2.68 477 54 A 2.65 498.455 A 2.33 423.5 56 A 2.61 499.4 57 A 2.67 515.4 58 A 2.37 449.6 59 A2.46 431.4 60 A 2.82 527 61 A 2.84

Example 27 Metalloenzyme Activity

A. Minimum Inhibitory Concentration (MIC) (C. albicans)

Compounds of the present disclosure were assessed for their ability toinhibit the growth of common strains of fungus, C. albicans using astandardized procedure (CLSI M27-A2).

Stock solutions of the test compounds and standards were prepared inDMSO at 1,600 μg/mL (C. albicans). Eleven, serial, one-half dilutions ofcompounds were prepared in 96-well plates in RPMI+MOPS. The assayconcentration ranges were 8-0.001 μg/mL (C. albicans). Cell suspensionsof C. albicans were prepared and added to each well at concentrations ofapproximately 3.7×10³ colony-forming-units per milliliter (cfu/mL). Alltesting was in duplicate. The inoculated plates were incubated forapproximately 48 h at 35±1° C. At the completion of incubation the wellsof each plate were evaluated visually for the presence of fungal growth.

For fluconazole and the test compounds, the MIC was the concentration atwhich growth was significantly reduced (about 50% reduction). Forvoriconazole the MIC was the concentration which reduced C. albicansgrowth by 50% (per CLS1, M27-A2). For QC purposes C. krusei isolate ATCC6258 (4.0×10³ cfu/mL) was included in the VOR assay. This isolate didnot exhibit trailing growth against voriconazole, therefore the MIC wasthe concentration at which growth was completely inhibited.

B. Inhibition of Liver Cytochrome P450 Enzymes

Solutions of each test compound were separately prepared atconcentrations of 20000, 6000, 2000, 600, 200, and 60 μM by serialdilution with DMSO:MeCN (50:50 v/v). The individual test compoundsolutions were then diluted 20-fold with DMSO:MeCN:deionized water(5:5:180 v/v/v) to concentrations of 1000, 300, 100, 30, 10, and 3 μM.Mixtures of isozyme inhibitors (sulfaphenazole, tranylcypromine, andketoconazole as specific inhibitors of isozymes 2C9, 2C19, and 3A4,respectively) were prepared containing each inhibitor at concentrationsof 6000, 2000, 600, 200, 60, 20, 6, and 2 μM by serial dilution withDMSO:ACN (50:50 v/v). The mixed inhibitor solutions were then diluted20-fold with DMSO:MeCN:deionized water (5:5:180 v/v/v) to concentrationsof 300, 100, 30, 10, 3, 1, 0.3, and 0.1 μM. The percent of organicsolvent attributable to the test compound or inhibitor mixture in thefinal reaction mixture was 2% v/v.

Pooled human liver microsome suspension (20 mg/mL) was diluted withphosphate buffer to obtain a 5 mg/mL suspension. A solution of NADPH wasprepared in phosphate buffer at a concentration of 5 mM. Separate stocksolutions of each substrate were prepared in DMSO:MeCN (50:50 v/v),mixed, and diluted in phosphate buffer to obtain a single solutioncontaining each substrate at five times its experimentally determinedK_(m) concentration. The percent of organic solvent attributable tosubstrate mixture in the final reaction mixture was 1% v/v.

Substrate solution and microsome suspension were combined in a 1:1volume ratio, mixed, and distributed to reaction wells of a PCR plate.Individual test compound or combined inhibitor solutions at eachconcentration were added to the wells and mixed by repetitiveaspirate-dispense cycles. For active controls, blank phosphate buffersolution was added in place of test compound solution. Reaction mixtureswere allowed to equilibrate at 37° C. for approximately two minutesbefore adding NADPH solution to initiate reaction, followed by pipettemixing of reaction mixture. Ten minutes after addition of NADPH, thereaction mixtures were quenched with cold acetonitrile. The samples weremixed by orbital shaking for approximately one minute and centrifuged at2900 RCF for ten minutes. A portion of the supernatant was analyzed bygradient reverse-phase HPLC with detection by electrospray ionizationtriple quadrupole mass spectrometry in the positive ion mode.

Data was fitted to sigmoid dose-response curves and the inhibitorypotency of each test compound was determined as its IC₅₀ value.

Results

Candida CYP2C9 Example MIC* IC50 CYP2C19 IC50 CYP3A4 IC50 9 0.016 13 1211 10 0.125 42 29 45 Fluconazole 0.5 29 8.2 8.0 Voriconazole 0.016 14 1513 Candida albicans MICs are in μg/mL; CYP IC50s are in μM.C. Minimum Inhibitory Concentration (MIC) (Septoria tritici)

Compounds of the present disclosure were assessed for their ability toinhibit the growth of a common strain of the fungal plant pathogenSeptoria tritici (ATCC 26517) using a procedure based on a Clinical andLaboratory Standards Institute (CLSI) microdilution assay protocol forfilamentous fungi.

Stock solutions of the test compounds and standards were prepared inDMSO at 6400 μg/mL. Each stock solution was used to prepare a 2-folddilution series ranging from 16 to 0.016 μg/mL (total of 11 compoundconcentrations) in RPMI-1640 (Roswell Park Memorial Institute) mediumcontaining 3-(N-morpholino)propanesulfonic acid (MOPS) buffer and 2%DMSO. A 100 μL aliquot of the dilutions was added to columns 1 (16 μg/mLcompound) through 11 (0.016 μg/mL compound) of a 96-well microtiterplate. This format was replicated in a second row of the microtiterplate. Thus, each microtiter plate could include 11 concentrations offour test or control compounds replicated twice. A 100 μL aliquot ofRPMI-1640/MOPS/2% DMSO medium was added to column 12 (no compoundcontrol) of the microtiter plate.

A fresh culture of S. tritici was used to prepare a solution ofapproximately 5×10⁴ colony-forming units per milliliter (cfu/mL) inRPMI/MOPS medium without DMSO. A 100 μL aliquot of this solution wasadded to all 96 wells in the microtiter plate. This results in finalconcentrations of each test or control compound of 8 μg/mL to 0.008μg/mL in 200 μL of RPMI/MOPS media containing 1% DMSO and approximately2.5×10⁴ cfu/mL of S. tritici. The assay plates were incubated at 22° C.for seven days in the dark without shaking. The MIC for each compoundwas visually determined as the concentration which resulted in 50%reduction in the growth of S. tritici in comparison to the control(column 12).

In each case of Table 3 the rating scale is as follows:

MIC (μg/mL Rating ≦0.5 A  >0.5-1.5 B  >1.5-4 C  >4 D Not tested E

TABLE 3 MIC Data for Compounds in Table 1 Patent Septoria Example#Rating  1 E  2 C  3 E  4 A  4(−) A  5 E  6 A  7 E  8(−) A  9(+) A 10(−)A 11 A 12 D 13 A 14 B 15 C 16 B 17 A 18 A 19 B 20 B 21 A 22 C 23 A 24 A25 A 26 A 27 A 28 A 29 A 30 C 31 C 32 A 34 A 35 C 36 C 37 A 38 A 39 A 40B 41 A 42 A 43 A 43(+) A 44 B 45 C 46 B 47 D 48 A 49 E 50 A 51 C 52 A 53C 54 C 55 C 56 B 57 A 58 C 59 B 60 C 61 BD. Evaluation of Fungicidal Activity vs. Leaf Rust (Causal AgentPuccinia recondita tritici=Puccinia triticina; Bayer code PUCCRT).

Wheat plants (variety Yuma) were grown from seed in a soil-lesspeat-based potting mixture (Metromix) until the seedlings had a fullyexpanded first leaf. Each pot contained 3-8 seedlings. These plants weresprayed until wet with the formulated test compounds. The compounds wereformulated at 50 ppm in 10 vol. % acetone plus 90 vol. % Triton X water(deionized water 99.99 wt %+0.01 wt % Triton X100), giving a “formulatedtest compound.” Formulated test compounds were applied to plants using aturn table sprayer fitted with two opposing air atomization nozzleswhich delivered approximately 1500 L/ha of spray volume. On thefollowing day, the leaves were inoculated with an aqueous sporesuspension of Puccinia recondita tritici and the plants were kept inhigh humidity overnight to permit spores to germinate and infect theleaf. The plants were then transferred to a greenhouse until diseasedeveloped on untreated control plants. Disease severity was evaluated7-9 days later, depending on the speed of disease development. Compounds4(−), 9(+), 11, 13, 18, 21, 25, 26, 27, 28, 32, 34, 37, 38, 39, 42, 43,48, 51, 52, 56, 57, and 59 were selected for testing against PUCCRT at50 ppm. Compounds that provided >80% disease control at 50 ppm included4(−), 9(+), 11, 13, 18, 21, 25, 26, 27, 28, 32, 34, 37, 38, 39, 42, 43,48, 52, 56, and 57.

Incorporation By Reference

The contents of all references (including literature references, issuedpatents, published patent applications, and co-pending patentapplications) cited throughout this application are hereby expresslyincorporated herein in their entireties by reference.

Equivalents

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents of the specificembodiments of the invention described herein. Such equivalents areintended to be encompassed by the following claims.

What is claimed:
 1. A method of treating a metalloenzyme-mediateddisease or disorder in or on a plant comprising contacting a compound ofFormula I with the plant or seeds, wherein:

MBG is optionally substituted tetrazoyl, optionally substitutedtriazolyl, optionally substituted oxazolyl, optionally substitutedpyrimidinyl, optionally substituted thiazolyl, or optionally substitutedpyrazolyl; R₁ is H, halo, alkyl or haloalkyl; R₂ is H, halo, alkyl orhaloalkyl; R₃ 1,1′-biphenyl substituted with 4′—OCH₂CF₃ or 4′-F, orheteroaryl, which may be optionally substituted with 1, 2 or 3independent R₅; R₄ is aryl, heteroaryl or cycloalkyl, optionallysubstituted with 0, 1, 2 or 3 independent R₆; each R₅ is independentlyH, halo, aryl optionally substituted 1, 2 or 3 independent R₆,heteroaryl, haloalkyl, haloalkoxy, cyano, nitro, alkyl, alkoxy, alkenyl,haloalkenyl, arylalkenyl, alkynyl, haloalkynyl, alklaryl, arylalkynyl,arylalkyl, cycloalklyl, halocycloalkyl, thioalkyl, SF₃, SF₆, SCN, SO₂R₇,C(O)alkyl, C(O)OH, C(O)Oalkyl; each R₆ is independently alkyl,thioalkyl, cyano, haloalkyl, hydroxyl, alkoxy, halo, haloalkoxy,—C(O)alkyl, —C(O)OH, —C(O)Oalkyl, SF₃, SF₆, SCN, SO₃H; and SO₂R₇; R₇ isindependently alkyl, aryl, substituted aryl, heteroaryl or substitutedheteroaryl; R₈ is H, —Si(R₉)₃, —P(O)(OH)₂, —CH₂—O—P(O)(OH)₂, or—C(O)alkyl optionally substituted with amino; R₉ is independently alkylor aryl; wherein R₃ is not optionally substituted monocyclic pyridyloptionally substituted with 1, 2 or 3 independent R₅; and themetalloenzyme-related disorder or disease is mediated by lanosteroldemethylase (CYP51).
 2. A method of inhibiting metalloenzyme activity ina microorganism on a plant comprising contacting a compound of Formula Iwith the plant or seeds, wherein:

MBG is optionally substituted tetrazoyl, optionally substitutedtriazolyl, optionally substituted oxazolyl, optionally substitutedpyrimidinyl, optionally substituted thiazolyl, or optionally substitutedpyrazolyl; R₁ is H, halo, alkyl or haloalkyl; R₂ is H, halo, alkyl orhaloalkyl; R₃ is 1,1′-biphenyl substituted with 4′—OCH₂CF₃ or 4′—F, orheteroaryl, which may be optionally substituted with 1, 2 or 3independent R₅; R₄ is aryl, heteroaryl or cycloalkyl, optionallysubstituted with 0, 1, 2or 3 independent R₆; each R₅ is independently H,halo, aryl optionally substituted 1, 2 or 3 independent R₆, heteroaryl,haloalkyl, haloalkoxy, cyano, nitro, alkyl, alkoxy, alkenyl,haloalkenyl, arylalkenyl, alkynyl, haloalkynyl, alklaryl, arylalkynyl,arylalkyl, cycloalklyl, halocycloalkyl, thioalkyl, SF₃, SF₆, SCN, SO₂R₇,C(O)alkyl, C(O)OH, C(O)Oalkyl; each R₆ is independently alkyl,thioalkyl, cyano, haloalkyl, hydroxyl, alkoxy, halo haloalkoxy,—C(O)alkyl, —C(O)OH, —C(O)Oalkyl, SF₃, SF₆, SCN, SO₃H; and SO₂R₇; R₇ isindependently alkyl, aryl, substituted aryl, heteroaryl or substitutedheteroaryl; R₈ is H, —Si(R₉)₃, —P(O)(OH)₂, —CH₂—O—P(O)(OH)₂, or—C(O)alkyl optionally substituted with amino; R₉ is independently alkylor aryl; wherein R₃ is not optionally substituted monocyclic pyridyloptionally substituted with 1, 2or 3 independent R₅; and wherein themetalloenzyme is lanosterol demethylase (CYP51).
 3. A method of treatinga fungal disease or disorder in or on a plant comprising contacting acompound of Formula I with the plant or seeds, wherein:

MBG is optionally substituted tetrazoyl, optionally substitutedtriazolyl, optionally substituted oxazolyl, optionally substitutedpyrimidinyl, optionally substituted thiazolyl, or optionally substitutedpyrazolyl; R₁ is H, halo, alkyl or haloalkyl; R₂ is H, halo, alkyl orhaloalkyl; R₃ is 1,1′-biphenyl substituted with 4′—OCH₂CF₃ or 4′—F, orheteroaryl, which may be optionally substituted with 1, 2 or 3independent R₅; R₄ is aryl, heteroaryl or cycloalkyl, optionallysubstituted with 0, 1, 2or 3 independent R₆; each R₅ is independently H,halo, aryl optionally substituted 1, 2 or 3 independent R₆, heteroaryl,haloalkyl, haloalkoxy, cyano, nitro, alkyl, alkoxy, alkenyl,haloalkenyl, arylalkenyl, alkynyl, haloalkynyl, alklaryl, arylalkynyl,arylalkyl, cycloalklyl, halocycloalkyl, thioalkyl, SF₃ , SF₆, SCN,SO₂R₇, C(O)alkyl, C(O)OH, C(O)Oalkyl; each R₆ is independently alkyl,thioalkyl, cyano, haloalkyl, hydroxyl, alkoxy, halo haloalkoxy,—C(O)alkyl, —C(O)OH, —C(O)Oalkyl, SF₃, SF₆, SCN, SO₃H; and SO₂R₇; R₇ isindependently alkyl, aryl, substituted aryl, heteroaryl or substitutedheteroaryl; R₈ is H, —Si(R₉)₃, —P(O)(OH)₂, —CH₂—O—P(O)(OH)₂, or—C(O)alkyl optionally substituted with amino; R₉ is independently alkylor aryl; and wherein R₃ is not optionally substituted monocyclic pyridyloptionally substituted with 1, 2or 3 independent R₅.
 4. A method oftreating fungal growth in or on a plant comprising contacting a compoundof Formula I with the plant or seeds, wherein:

MBG is optionally substituted tetrazoyl, optionally substitutedtriazolyl, optionally substituted oxazolyl, optionally substitutedpyrimidinyl, optionally substituted thiazolyl, or optionally substitutedpyrazolyl; R₁ is H, halo, alkyl or haloalkyl; R₂ is H, halo, alkyl orhaloalkyl; R₃ is 1,1′-biphenyl substituted with 4′—OCH₂CF₃ or 4′—F, orheteroaryl, which may be optionally substituted with 1, 2 or 3independent R₅; R₄ is aryl, heteroaryl or cycloalkyl, optionallysubstituted with 0, 1, 2or 3 independent R₆; each R₅ is independently H,halo, aryl optionally substituted 1, 2 or 3 independent R₆, heteroaryl,haloalkyl, haloalkoxy, cyano, nitro, alkyl, alkoxy, alkenyl,haloalkenyl, arylalkenyl, alkynyl, haloalkynyl, alklaryl, arylalkynyl,arylalkyl, cycloalklyl, halocycloalkyl, thioalkyl, SF₃ , SF₆, SCN,SO₂R₇, C(O)alkyl, C(O)OH, C(O)Oalkyl; each R₆ is independently alkyl,thioalkyl, cyano, haloalkyl, hydroxyl, alkoxy, halo haloalkoxy,—C(O)alkyl, —C(O)OH, —C(O)Oalkyl, SF₃, SF₆, SCN, SO₃H; and SO₂R₇; R₇ isindependently alkyl, aryl, substituted aryl, heteroaryl or substitutedheteroaryl; R₈ is H, —Si(R₉)₃, —P(O)(OH)₂, —CH₂—O—P(O)(OH)₂, or—C(O)alkyl optionally substituted with amino; R₉ is independently alkylor aryl; and wherein R₃ is not optionally substituted monocyclic pyridyloptionally substituted with 1, 2 or 3 independent R₅.
 5. A method ofinhibiting microorganisms in or on a plant comprising contacting acompound of Formula I with the plant or seeds, wherein:

MBG is optionally substituted tetrazoyl, optionally substitutedtriazolyl, optionally substituted oxazolyl, optionally substitutedpyrimidinyl, optionally substituted thiazolyl, or optionally substitutedpyrazolyl; R₁ is H, halo, alkyl or haloalkyl; R₂ is H, halo, alkyl orhaloalkyl; R₃ is 1,1′-biphenyl substituted with 4′—OCH₂CF₃ or 4′—F, orheteroaryl, which may be optionally substituted with 1, 2 or 3independent R₅; R₄ is aryl, heteroaryl or cycloalkyl, optionallysubstituted with 0, 1, 2 or 3 independent R_(6;) each R₅ isindependently H, halo, aryl optionally substituted 1, 2 or 3 independentR₆, heteroaryl, haloalkyl, haloalkoxy, cyano, nitro, alkyl, alkoxy,alkenyl, haloalkenyl, arylalkenyl, alkynyl, haloalkynyl, alklaryl,arylalkynyl, arylalkyl, cycloalklyl, halocycloalkyl, thioalkyl, SF₃ ,SF₆, SCN, SO₂R₇, C(O)alkyl, C(O)OH, C(O)Oalkyl; each R₆ is independentlyalkyl, thioalkyl, cyano, haloalkyl, hydroxyl, alkoxy, halo haloalkoxy,—C(O)alkyl, —C(O)OH, —C(O)Oalkyl, SF₃, SF₆, SCN, SO₃H; and SO₂R₇; R₇ isindependently alkyl, aryl, substituted aryl, heteroaryl or substitutedheteroaryl; R₈ is H, —Si(R₉)₃, —P(O)(OH)₂, —CH₂—O—P(O)(OH)₂, or—C(O)alkyl optionally substituted with amino; R₉ is independently alkylor aryl; and wherein R₃ is not optionally substituted monocyclic pyridyloptionally substituted with 1, 2 or 3 independent R₅; and wherein themicroorganism belongs to at least one genera selected from Blumeria,Podospaera, Sphaerotheca, Unicinula, Erysiphe, Puccinia, Phakopspora,Gymnosporangium, Hemileia, Uromyces, Alternia, Cercospora, Cladosporium,Cochliobolus, Colletotrichum, Magnaporthe, Mycosphaerella,Phaeosphaeria, Pyrenophora, Ramularia, Rhyncosporium, Septoria,Venturia, Ustilago, Aspergillus, Penicillium, Drechslera, Fusarium,Botrytis, Gibberella, Rhizoctonia, Pseudocercosporella, Sclerotinia,Helminithosporium, Stagonospora, Exserohilum, and Pyricularia.
 6. Acomposition comprising a compound of Formula I or salt thereof, wherein:

MBG is optionally substituted tetrazoyl, optionally substitutedtriazolyl, optionally substituted oxazolyl, optionally substitutedpyrimidinyl, optionally substituted thiazolyl, or optionally substitutedpyrazolyl; R₁ is H, halo, alkyl or haloalkyl; R₂ is H, halo, alkyl orhaloalkyl; R₃ is 1,1′-biphenyl substituted with 4′—OCH₂CF₃ or 4′—F, orheteroaryl, which may be optionally substituted with 1, 2 or 3independent R₅; R₄ is aryl, heteroaryl or cycloalkyl, optionallysubstituted with 0, 1, 2 or 3 independent R₆; each R₅ is independentlyH, halo, aryl optionally substituted 1, 2 or 3 independent R₆,heteroaryl, haloalkyl, haloalkoxy, cyano, nitro, alkyl, alkoxy, alkenyl,haloalkenyl, arylalkenyl, alkynyl, haloalkynyl, alklaryl, arylalkynyl,arylalkyl, cycloalklyl, halocycloalkyl, thioalkyl, SF₃ , SF₆, SCN,SO₂R₇, C(O)alkyl, C(O)OH, C(O)Oalkyl; each R₆ is independently alkyl,thioalkyl, cyano, haloalkyl, hydroxyl, alkoxy, halo haloalkoxy,—C(O)alkyl, —C(O)OH, —C(O)Oalkyl, SF₃, SF₆, SCN, SO₃H; and SO₂R₇; R₇ isindependently alkyl, aryl, substituted aryl, heteroaryl or substitutedheteroaryl; R₈ is H, —Si(R₉)₃, —P(O)(OH)₂, —CH₂—O—P(O)(OH)₂, or—C(O)alkyl optionally substituted with amino; R₉ is independently alkylor aryl; and wherein R₃ is not optionally substituted monocyclic pyridyloptionally substituted with 1, 2 or 3 independent R₅, said compositionfurther comprising an azole fungicide selected from epoxiconazole,tebuconazole, fluquinconazole, flutriafol, metaconazole, myclobutanil,cycproconazole, prothioconazole and propiconazole.
 7. A compositioncomprising a compound of Formula I, or salt thereof, wherein:

MBG is optionally substituted tetrazoyl, optionally substitutedtriazolyl, optionally substituted oxazolyl, optionally substitutedpyrimidinyl, optionally substituted thiazolyl, or optionally substitutedpyrazolyl; R₁ is H, halo, alkyl or haloalkyl; R₂ is H, halo, alkyl orhaloalkyl; R₃ is 1,1′-biphenyl substituted with 4′—OCH₂CF₃ or 4′—F, orheteroaryl, which may be optionally substituted with 1, 2 or 3independent R₅; R₄ is aryl, heteroaryl or cycloalkyl, optionallysubstituted with 0, 1, 2or 3 independent R₆; each R₅ is independently H,halo, aryl optionally substituted 1, 2 or 3 independent R₆, heteroaryl,haloalkyl, haloalkoxy, cyano, nitro, alkyl, alkoxy, alkenyl,haloalkenyl, arylalkenyl, alkynyl, haloalkynyl, alklaryl, arylalkynyl,arylalkyl, cycloalklyl, halocycloalkyl, thioalkyl, SF₃ , SF₆, SCN,SO₂R₇, C(O)alkyl, C(O)OH, C(O)Oalkyl; each R₆ is independently alkyl,thioalkyl, cyano, haloalkyl, hydroxyl, alkoxy, halo haloalkoxy,—C(O)alkyl, —C(O)OH, —C(O)Oalkyl, SF₃, SF₆, SCN, SO₃H; and SO₂R₇; R₇ isindependently alkyl, aryl, substituted aryl, heteroaryl or substitutedheteroaryl; R₈ is H, —Si(R₉)₃, —P(O)(OH)₂, —CH₂—O—P(O)(OH)₂, or—C(O)alkyl optionally substituted with amino; R₉ is independently alkylor aryl; and wherein R₃ is not optionally substituted monocyclic pyridyloptionally substituted with 1, 2 or 3 independent R₅, said compositionfurther comprising a strobilurin fungicide from the grouptrifloxystrobin, pyraclostrobin, orysastrobin, fluoxastrobin andazoxystrobin.
 8. A method of inhibiting microorganisms in or on a plantcomprising contacting a compound of Formula I with the plant or seeds,wherein:

MBG is optionally substituted tetrazoyl, optionally substitutedtriazolyl, optionally substituted oxazolyl, optionally substitutedpyrimidinyl, optionally substituted thiazolyl, or optionally substitutedpyrazolyl; R₁ is H, halo, alkyl or haloalkyl; R₂ is H, halo, alkyl orhaloalkyl; R₃ is 1,1′-biphenyl substituted with 4′—OCH₂CF₃ or 4′—F, orheteroaryl, which may be optionally substituted with 1, 2 or 3independent R₅; R₄ is aryl, heteroaryl or cycloalkyl, optionallysubstituted with 0, 1, 2or 3 independent R₆; each R₅ is independently H,halo, aryl optionally substituted 1, 2 or 3 independent R₆, heteroaryl,haloalkyl, haloalkoxy, cyano, nitro, alkyl, alkoxy, alkenyl,haloalkenyl, arylalkenyl, alkynyl, haloalkynyl, alklaryl, arylalkynyl,arylalkyl, cycloalklyl, halocycloalkyl, thioalkyl, SF₃, SF₆, SCN, SO₂R₇,C(O)alkyl, C(O)OH, C(O)Oalkyl; each R₆ is independently alkyl,thioalkyl, cyano, haloalkyl, hydroxyl, alkoxy, halo haloalkoxy,—C(O)alkyl, —C(O)OH, —C(O)Oalkyl, SF₃, SF₆, SCN, SO₃H; and SO₂R₇; R₇ isindependently alkyl, aryl, substituted aryl, heteroaryl or substitutedheteroaryl; R₈ is H, —Si(R₉)₃, —P(O)(OH)₂, —CH₂—O—P(O)(OH)₂, or—C(O)alkyl optionally substituted with amino; R₉ is independently alkylor aryl; and wherein R₃ is not optionally substituted monocyclic pyridyloptionally substituted with 1, 2 or 3 independent R₅; and wherein themicroorganism is selected from the group consisting of Venturiainaequalis, Septoria tritici, Cercospora beticola, Cercosporaarachidicola, Colletotrichum lagenarium, Puccinia graminis f. sp.tritici, Puccinia recondite tritici, Uncinula necoator, Blumeriagrammis, and Mycosphaerella fijiensis.
 9. The method of claim 3, whereinthe fungal disease or disorder is selected from the group consisting ofapple scab, speckled leaf blotch of wheat, leaf spot of sugarbeets, leafspot of peanut, cucumber anthracnose, wheat leaf rust, grape powderymildew, wheat powdery mildew, and black sigatoka.
 10. The method ofclaim 4, wherein R₁ is fluoro.
 11. The method of claim 4, wherein R₂ isfluoro.
 12. The method of claim 4, wherein R₁ and R₂ are fluoro.
 13. Themethod of claim 4, wherein R₄ is phenyl optionally substituted with 0,1, 2 or 3 independent R₆.
 14. The method of claim 4, wherein R₄ isphenyl optionally substituted with 0, 1, 2 or 3 independent halo. 15.The method of claim 4, wherein R₄ is phenyl optionally substituted with0, 1, 2 or 3 independent fluoro.
 16. The method of claim 4, wherein R₁is 2,4-difluorophenyl.
 17. The method of claim 4, wherein R₅ is halo.18. The method of claim 4, wherein R₃ is heteroaryl optionallysubstituted with 1, 2 or 3 independent R₅; wherein at least one R₅ ishalo.
 19. The method of claim 4, wherein: R₁ is fluoro; R₂ is fluoro; R₄is 2,4-difluorophenyl; and R₃ is heteroaryl other than 2-pyridyl,substituted with 1, 2 or 3 independent R₅.
 20. The method of claim 4,wherein: R₁ is fluoro; R₂ is fluoro; R₄ is 2,4-difluorophenyl; and R₃ isbicyclic heteroaryl substituted with 1, 2 or 3 independent R₅.
 21. Themethod of claim 4, wherein: R₃ is 2-quinolinyl substituted with 1, 2 or3 independent R₅.
 22. The method of claim 4, wherein the compound is oneof:1-(5-Chlorothiophen-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(1);1-(4-Bromothiazol-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(2);4-(2-(2-(2,4-Difluorophenyl)-1,1-difluoro-2-hydroxy-3-(1H-tetrazol-1-yl)propyl)thiazol-4-yl)benzonitrile(3);1-(6-Chloroquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(4);2-(2,4-Difluorophenyl)-1,1-difluoro-1-(quinolin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(5);1-(Benzo[d]thiazol-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(6);2-(2,4-Difluorophenyl)-1,1-difluoro-1-(pyrimidin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(7);2-(4-Chloro-2-fluorophenyl)-1-(6-chloroquinolin-2-yl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(8);1-(6-Bromoquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(9);1-(6-Chloroquinoxalin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(10);1-(6-Chlorobenzo[d]thiazol-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(11);2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(thiazol-2-yl)propan-2-ol(12);1-(5-Bromothiophen-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(13);2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(thiophen-2-yl)propan-2-ol(14);1-(6-Chloroquinolin-2-yl)-1,1-difluoro-2-(4-methoxyphenyl)-3-(1H-tetrazol-1-yl)propan-2-ol(15);1-(6-Chloroquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(2H-tetrazol-2-yl)propan-2-ol(16);2-(2,4-Difluorophenyl)-1,1-difluoro-1-(6-fluoroquinolin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(17);2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-(trifluoromethyl)quinolin-2-yl)propan-2-ol(18);2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-(2,2,2-trifluoroethoxy)quinolin-2-yl)propan-2-ol(19);1-(6-Chloroquinolin-2-yl)-1,1-difluoro-2-(2-fluoro-4-(trifluoromethyl)phenyl)-3-(1H-tetrazol-1-yl)propan-2-ol(20);2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-(trifluoromethoxy)quinolin-2-yl)propan-2-ol(21);2-(2-Chloro-4-(trifluoromethyl)phenyl)-1-(6-chloroquinolin-2-yl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(22);1-(6-Chloroquinolin-2-yl)-2-(3,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(23);2-(2-(2,4-Difluorophenyl)-1,1-difluoro-2-hydroxy-3-(1H-tetrazol-1-yl)propyl)quinoline-6-carbonitrile(24);1-(6-(Difluoromethyl)quinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(25);2-(2,4-Difluorophenyl)-1,1-difluoro-1-(6-methylquinolin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(26);1-(6-Bromobenzo[d]thiazol-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(27);1-(6-Chloroquinolin-2-yl)-2-(2,5-difluorophenyl)-1,1-difluoro-3-(2H-tetrazol-2-yl)propan-2-ol(28);1-(5,6-Dichloroquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(29);2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(2,2,2-trifluoroethoxy)quinolin-2-yl)propan-2-ol(30);1-(5-Chloroquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(31);1-(6-Chloroquinolin-2-yl)-1,1-difluoro-2-(4-fluorophenyl)-3-(1H-tetrazol-1-yl)propan-2-ol(32);1-(6-Cloroquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-1,2,4-triazol-1-yl)propan-2-ol(33);2-(4-Chloro-2-fluorophenyl)-1-(6-chloroquinoxalin-2-yl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(34);1-(6-Chloroquinolin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(4H-1,2,4-triazol-4-yl)propan-2-ol(35);1-(7-Chloroisoquinolin-3-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(37);1-(6-Bromoquinoxalin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(38);1-(5-(4-(Difluoromethoxy)phenyl)pyrazin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(39);1-(5-(4-Chlorophenyl)pyrazin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(40);2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyrazin-2-yl)propan-2-ol(41);2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(trifluoromethoxy)phenyl)pyrazin-2-yl)propan-2-ol(42);1-(5-(4-Bromophenyl)pyrazin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(43);2-(2,4-Difluorophenyl)-1-(5-(3,4-difluorophenyl)pyrazin-2-yl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(44);1,1-Difluoro-2-(4-fluorophenyl)-3-(1H-tetrazol-1-yl)-1-(5-(4-(trifluoromethoxy)phenyl)pyrazin-2-yl)propan-2-ol(45);2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(4-fluorophenyl)pyrazin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(46);2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(4-fluorophenyl)pyrazin-2-yl)-3-(2H-tetrazol-2-yl)propan-2-ol(47);2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(4-methoxyphenyl)pyrazin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(48);1-(5-Chloropyrazin-2-yl)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)propan-2-ol(49);2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-((4-fluorophenyl)ethynyl)pyrazin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(50);2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-((4-fluorophenyl)ethynyl)pyrazin-2-yl)-3-(2H-tetrazol-2-yl)propan-2-ol(51);2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(4-fluorophenethyl)pyrazin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(52);2-(2,4-Difluorophenyl)-1,1-difluoro-1-(5-(4-fluorophenethyl)pyrazin-2-yl)-3-(2H-tetrazol-2-yl)propan-2-ol(53);2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-(trifluoromethoxy)quinoxalin-2-yl)propan-2-ol(54);2-(2,4-Difluorophenyl)-1,1-difluoro-1-(6-fluoroquinoxalin-2-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(55);2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-(4-(trifluoromethyl)phenyl)pyridazin-3-yl)propan-2-ol(56);2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-(4-(trifluoromethoxy)phenyl)pyridazin-3-yl)propan-2-ol(57);2-(2,4-Difluorophenyl)-1,1-difluoro-1-(6-(4-fluorophenyl)pyridazin-3-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(58);2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(6-vinylquinoxalin-2-yl)propan-2-ol(59);2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(4′-(2,2,2-trifluoroethoxy)-[1,1′-biphenyl]-4-yl)propan-2-ol(60);2-(2,4-Difluorophenyl)-1,1-difluoro-1-(4′-fluoro-[1,1′-biphenyl]-4-yl)-3-(1H-tetrazol-1-yl)propan-2-ol(61).